subject:(recombinant protein)

University of Manchester

1. French, Joseph. Towards an understanding of the burden of recombinant protein production.

Degree: 2016, University of Manchester

URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:294150

► Recombinant protein technologies have emerged as important tools for the production of proteins with industrial, academic and biopharmaceutical applications. However, current process development for a… (more)

▼ Recombinant protein technologies have emerged as important tools for the production of proteins with industrial, academic and biopharmaceutical applications. However, current process development for a target protein is hindered by the burden recombinant protein expression places on the host system, with the level of this negative effect and the ideal production conditions varying from protein to protein. As a result current process optimisation relies on trial and error to determine the optimal set up for a given target protein. The work presented here will examine two mechanisms by which this burden acts on the cell, contributing towards making the overall burden effect and the optimal process conditions more predictable.Flux Balance Analysis was used to examine the effect of amino acid supplementation on the metabolic cost of a recombinant protein to predict which supplements would improve the efficiency of production, predictions supported elsewhere in the literature. However, experimental validation in batch and fed batch cultures for the production of human Granulocyte Colony Stimulating Factor demonstrated that these supplementation strategies do not lead to an increase in yield or performance. The results from the computational modelling alongside similar studies in the literature suggest that the more important factor may be optimisation for better growth generally rather than targeted attempts based on the protein composition.The sensitivity of native E. coli proteins to a loss of chaperone activity was predicted using the solubility data of the eSol database, identifying rrmJ as a protein of interest. The possible significance of rrmJ for chaperone saturation was examined alongside examining the effect of recombinant protein solubility using Green Fluorescent Protein (GFP) and its mutant GFP_A which have differing solubility. However, neither an effect through rrmJ nor a negative effect of recombinant protein solubility on growth was identified. Kinetic modelling for a mechanistic examination of the chaperone network suggests this is because the poorly folding protein is preferentially shifted to the insoluble fraction while the better performing proteins, i.e. the native proteins, are relatively unperturbed despite saturation of the chaperones.Overall the study was not able to make these areas more predictable. However, the observations made within this study contribute to an improvement in our understanding of two key mechanisms of interest in the field. Of particular interest is the identification of two logical hypotheses within the literature to be, at least in the cases tested, false. Advisors/Committee Members: WARWICKER, JIM J, DICKSON, ALAN AJ, SNOEP, JACOB JL, Warwicker, Jim, Dickson, Alan, Westerhoff, Hans, Snoep, Jacob.

Subjects/Keywords: recombinant protein; escherichia coli; burden

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APA (6^th Edition):

French, J. (2016). Towards an understanding of the burden of recombinant protein production. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:294150

Chicago Manual of Style (16^th Edition):

French, Joseph. “Towards an understanding of the burden of recombinant protein production.” 2016. Doctoral Dissertation, University of Manchester. Accessed January 24, 2021. http://www.manchester.ac.uk/escholar/uk-ac-man-scw:294150.

MLA Handbook (7^th Edition):

French, Joseph. “Towards an understanding of the burden of recombinant protein production.” 2016. Web. 24 Jan 2021.

Vancouver:

French J. Towards an understanding of the burden of recombinant protein production. [Internet] [Doctoral dissertation]. University of Manchester; 2016. [cited 2021 Jan 24]. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:294150.

Council of Science Editors:

French J. Towards an understanding of the burden of recombinant protein production. [Doctoral Dissertation]. University of Manchester; 2016. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:294150

Purdue University

2. Brennan, Mary Jane. Design and Characterization of Biomimetic Adhesive Materials.

Degree: PhD, Chemical Engineering, 2015, Purdue University

URL: https://docs.lib.purdue.edu/open_access_dissertations/1340

► When we engineer new materials, nature provides us with a wealth of inspiration, often in the form of proteins. The blue mussel Mytilus edulis and… (more)

Subjects/Keywords: adhesion; bioinspired; elastomeric; protein; recombinant

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APA (6^th Edition):

Brennan, M. J. (2015). Design and Characterization of Biomimetic Adhesive Materials. (Doctoral Dissertation). Purdue University. Retrieved from https://docs.lib.purdue.edu/open_access_dissertations/1340

Chicago Manual of Style (16^th Edition):

Brennan, Mary Jane. “Design and Characterization of Biomimetic Adhesive Materials.” 2015. Doctoral Dissertation, Purdue University. Accessed January 24, 2021. https://docs.lib.purdue.edu/open_access_dissertations/1340.

MLA Handbook (7^th Edition):

Brennan, Mary Jane. “Design and Characterization of Biomimetic Adhesive Materials.” 2015. Web. 24 Jan 2021.

Vancouver:

Brennan MJ. Design and Characterization of Biomimetic Adhesive Materials. [Internet] [Doctoral dissertation]. Purdue University; 2015. [cited 2021 Jan 24]. Available from: https://docs.lib.purdue.edu/open_access_dissertations/1340.

Council of Science Editors:

Brennan MJ. Design and Characterization of Biomimetic Adhesive Materials. [Doctoral Dissertation]. Purdue University; 2015. Available from: https://docs.lib.purdue.edu/open_access_dissertations/1340

Stellenbosch University

3. De Villiers, Ann-Marie. Production and glycosylation of a recombinant protein from Chinese hamster ovary (CHO) cells.

Degree: MScEng, Process Engineering, 2012, Stellenbosch University

URL: http://hdl.handle.net/10019.1/71663

► ENGLISH ABSTRACT: Recombinant glycoproteins are important biopharmaceuticals, providing solutions for numerous previously untreatable illnesses, in everything from cancer to infertility. Most recombinant biopharmaceuticals are produced… (more)

▼ ENGLISH ABSTRACT: Recombinant glycoproteins are important biopharmaceuticals, providing solutions for numerous previously untreatable illnesses, in everything from cancer to infertility. Most recombinant biopharmaceuticals are produced in mammalian cells due to their ability to provide the correct post-translational processing for use in humans. The post-translation processing influences many of the protein’s properties including pharmacokinetics, bioactivity, secretion, half-life, solubility, recognition and antigenicity. The aim of this thesis is to further study the upstream production of a glycosylated recombinant protein produced by Chinese hamster ovary (CHO) cells on production scale within the confines of an existing process. The process in question uses adherent CHO cells to produce a glycosylated recombinant hormone. As with most recombinant protein production processes, this process has two sections to the upstream production: a seed train to grow enough cells to inoculate production, and a production section, which focuses on the production of a recombinant protein. The seed train is predominantly conducted in roller bottles, while the production section takes place in perfusion bioreactors, where the cells are attached to microcarriers, with spin-filters for cell retention. The whole process uses medium with serum. There are two process challenges regarding an existing recombinant-protein production process: 1. The gradual increase, over the past several campaigns, of the final population doubling level of the cells (which must remain within certain specified limits) at the end of the seed train. 2. The low glycosylation levels of the product seen in certain campaigns, which meant that a certain number of final product batches were below the specified acceptable glycosylation limits. Following a literature survey several controlled process variables were chosen for investigation and hypotheses made on their effect on the seed train or glycosylation. To investigate their effect on the PDL and cell growth in the seed train: - Medium volume: decreasing the medium volume will yield a lower PDL due to slower cell growth caused by lower glucose availability. - Seeding density: if cells obtain confluence by the time they are harvested, decreasing the seeding density will yield a higher PDL. - Cultivation temperature: decreasing the temperature ought to decrease the growth rate. - Medium feed temperature: there will be no significant difference to the cell culture when pre-heated or cold medium is used. Aeration: using vent caps will increase the oxygen content of the medium in the roller bottles and the cell growth, yielding a higher PDL. To investigate their effect on glycosylation during production: - pH: better glycosylation will be seen at pH 6.9, than at pH 6.7. - Perfusion rate: a higher perfusion rate will lead to better glycosylation due to increased glucose and glutamine concentrations. In the seed train, the only factor that significantly influenced the final PDL was the seeding density.… Advisors/Committee Members: Gorgens, Johann F., Stellenbosch University. Faculty of Engineering. Dept. of Process Engineering..

Subjects/Keywords: Process engineering; Recombinant glycoproteins; Biopharmaceuticals; Glycosylated recombinant protein – Production

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APA (6^th Edition):

De Villiers, A. (2012). Production and glycosylation of a recombinant protein from Chinese hamster ovary (CHO) cells. (Masters Thesis). Stellenbosch University. Retrieved from http://hdl.handle.net/10019.1/71663

Chicago Manual of Style (16^th Edition):

De Villiers, Ann-Marie. “Production and glycosylation of a recombinant protein from Chinese hamster ovary (CHO) cells.” 2012. Masters Thesis, Stellenbosch University. Accessed January 24, 2021. http://hdl.handle.net/10019.1/71663.

MLA Handbook (7^th Edition):

De Villiers, Ann-Marie. “Production and glycosylation of a recombinant protein from Chinese hamster ovary (CHO) cells.” 2012. Web. 24 Jan 2021.

Vancouver:

De Villiers A. Production and glycosylation of a recombinant protein from Chinese hamster ovary (CHO) cells. [Internet] [Masters thesis]. Stellenbosch University; 2012. [cited 2021 Jan 24]. Available from: http://hdl.handle.net/10019.1/71663.

Council of Science Editors:

De Villiers A. Production and glycosylation of a recombinant protein from Chinese hamster ovary (CHO) cells. [Masters Thesis]. Stellenbosch University; 2012. Available from: http://hdl.handle.net/10019.1/71663

University of Ottawa

4. Podrebarac, James. Development of Recombinant Human Collagen Type I and Type III Injectable Hydrogels for Cardiac Therapy .

Degree: 2017, University of Ottawa

URL: http://hdl.handle.net/10393/36038

► Functional biomaterials are being developed as scaffolds to support endogenous cells and to promote the regeneration of ischemic tissue. The aim for this study was… (more)

Subjects/Keywords: Biomaterials; Cardiovascular disease; Collagen; Injectable; Recombinant protein

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APA (6^th Edition):

Podrebarac, J. (2017). Development of Recombinant Human Collagen Type I and Type III Injectable Hydrogels for Cardiac Therapy . (Thesis). University of Ottawa. Retrieved from http://hdl.handle.net/10393/36038

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Podrebarac, James. “Development of Recombinant Human Collagen Type I and Type III Injectable Hydrogels for Cardiac Therapy .” 2017. Thesis, University of Ottawa. Accessed January 24, 2021. http://hdl.handle.net/10393/36038.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Podrebarac, James. “Development of Recombinant Human Collagen Type I and Type III Injectable Hydrogels for Cardiac Therapy .” 2017. Web. 24 Jan 2021.

Vancouver:

Podrebarac J. Development of Recombinant Human Collagen Type I and Type III Injectable Hydrogels for Cardiac Therapy . [Internet] [Thesis]. University of Ottawa; 2017. [cited 2021 Jan 24]. Available from: http://hdl.handle.net/10393/36038.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Podrebarac J. Development of Recombinant Human Collagen Type I and Type III Injectable Hydrogels for Cardiac Therapy . [Thesis]. University of Ottawa; 2017. Available from: http://hdl.handle.net/10393/36038

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Queensland University of Technology

5. Tan, Yu Pei. The development of Lactococcus lactis as an antimicrobial agent.

Degree: 2010, Queensland University of Technology

URL: https://eprints.qut.edu.au/39143/

► Non-pathogenic lactic acid bacteria are economically important Gram-positive bacteria used extensively in the food industry. Due to their “generally regarded as safe” status, certain species… (more)

▼ Non-pathogenic lactic acid bacteria are economically important Gram-positive bacteria used extensively in the food industry. Due to their “generally regarded as safe” status, certain species from the genera Lactobacillus and Lactococcus are also considered desirable as candidates for the production and secretion of recombinant proteins, particular those with therapeutic applications. The hypothesis examined by this thesis is that Lactococcus lactis can be modified to be an effective antimicrobial agent. Therefore, the aims of this thesis were to investigate the optimisation of the expression, secretion and/or activities of potential heterologous antimicrobial proteins by the model lactic acid bacterium, Lactococcus lactis subsp. cremoris MG1363. L. lactis strains were engineered to express and secrete the recombinant CyuC surface protein from Lactobacillus reuteri BR11, and a fusion protein consisting of CyuC and lysostaphin using the Sep promoter and secretion signal. CyuC has been characterised as a cystine-binding protein, but has also been demonstrated to have fibronectin binding activity. Lysostaphin is a bacteriolytic enzyme with specific activity against the Gram-positive pathogen, Staphylococcus aureus. These modified L. lactis strains were then investigated to see if they had the ability to inhibit the adhesion of S. aureus to host extracellular matrix (ECM) proteins. It was observed that the cell extracts of the L. lactis strain with the vector only (pGhost9:ISS1) was able to inhibit the adhesion of S. aureus to fibronectin, whilst the cell extracts of the L. lactis strain expressing lysostaphin was able to inhibit adhesion to keratin. Finally, this thesis has identified specific lactococcal genes that affect the secretion of lysostaphin through the use of random transposon mutagenesis. Ten mutants with higher lysostaphin activity contained insertions in four different genes encoding: (i) an uncharacterised putative transmembrane protein (llmg_0609), (ii) an enzyme catalysing the first step in peptidoglycan biosynthesis (murA2), (iii) a homolog of the oxidative defence regulator (trmA), and (iv) an uncharacterised putative enzyme involved in ubiquinone biosynthesis (llmg_2148). The higher lysostaphin activity observed in these mutants was found to be due to higher amounts of lysostaphin being secreted. The findings of this thesis contribute to the development of this organism as an antimicrobial agent and also to our understanding of L. lactis genetics.

Subjects/Keywords: Lactococcus; recombinant protein secretion; lysostaphin; ECM proteins

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Tan, Y. P. (2010). The development of Lactococcus lactis as an antimicrobial agent. (Thesis). Queensland University of Technology. Retrieved from https://eprints.qut.edu.au/39143/

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Tan, Yu Pei. “The development of Lactococcus lactis as an antimicrobial agent.” 2010. Thesis, Queensland University of Technology. Accessed January 24, 2021. https://eprints.qut.edu.au/39143/.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Tan, Yu Pei. “The development of Lactococcus lactis as an antimicrobial agent.” 2010. Web. 24 Jan 2021.

Vancouver:

Tan YP. The development of Lactococcus lactis as an antimicrobial agent. [Internet] [Thesis]. Queensland University of Technology; 2010. [cited 2021 Jan 24]. Available from: https://eprints.qut.edu.au/39143/.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Tan YP. The development of Lactococcus lactis as an antimicrobial agent. [Thesis]. Queensland University of Technology; 2010. Available from: https://eprints.qut.edu.au/39143/

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Manchester

6. French, Joseph. Towards an understanding of the burden of recombinant protein production.

Degree: PhD, 2016, University of Manchester

URL: https://www.research.manchester.ac.uk/portal/en/theses/towards-an-understanding-of-the-burden-of-recombinant-protein-production(4c331186-592a-4984-88de-70a2a27c9fb9).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.679990

► Recombinant protein technologies have emerged as important tools for the production of proteins with industrial, academic and biopharmaceutical applications. However, current process development for a… (more)

▼ Recombinant protein technologies have emerged as important tools for the production of proteins with industrial, academic and biopharmaceutical applications. However, current process development for a target protein is hindered by the burden recombinant protein expression places on the host system, with the level of this negative effect and the ideal production conditions varying from protein to protein. As a result current process optimisation relies on trial and error to determine the optimal set up for a given target protein. The work presented here will examine two mechanisms by which this burden acts on the cell, contributing towards making the overall burden effect and the optimal process conditions more predictable. Flux Balance Analysis was used to examine the effect of amino acid supplementation on the metabolic cost of a recombinant protein to predict which supplements would improve the efficiency of production, predictions supported elsewhere in the literature. However, experimental validation in batch and fed batch cultures for the production of human Granulocyte Colony Stimulating Factor demonstrated that these supplementation strategies do not lead to an increase in yield or performance. The results from the computational modelling alongside similar studies in the literature suggest that the more important factor may be optimisation for better growth generally rather than targeted attempts based on the protein composition. The sensitivity of native E. coli proteins to a loss of chaperone activity was predicted using the solubility data of the eSol database, identifying rrmJ as a protein of interest. The possible significance of rrmJ for chaperone saturation was examined alongside examining the effect of recombinant protein solubility using Green Fluorescent Protein (GFP) and its mutant GFP_A which have differing solubility. However, neither an effect through rrmJ nor a negative effect of recombinant protein solubility on growth was identified. Kinetic modelling for a mechanistic examination of the chaperone network suggests this is because the poorly folding protein is preferentially shifted to the insoluble fraction while the better performing proteins, i.e. the native proteins, are relatively unperturbed despite saturation of the chaperones. Overall the study was not able to make these areas more predictable. However, the observations made within this study contribute to an improvement in our understanding of two key mechanisms of interest in the field. Of particular interest is the identification of two logical hypotheses within the literature to be, at least in the cases tested, false.

Subjects/Keywords: 572; recombinant protein; escherichia coli; burden

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

French, J. (2016). Towards an understanding of the burden of recombinant protein production. (Doctoral Dissertation). University of Manchester. Retrieved from https://www.research.manchester.ac.uk/portal/en/theses/towards-an-understanding-of-the-burden-of-recombinant-protein-production(4c331186-592a-4984-88de-70a2a27c9fb9).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.679990

Chicago Manual of Style (16^th Edition):

French, Joseph. “Towards an understanding of the burden of recombinant protein production.” 2016. Doctoral Dissertation, University of Manchester. Accessed January 24, 2021. https://www.research.manchester.ac.uk/portal/en/theses/towards-an-understanding-of-the-burden-of-recombinant-protein-production(4c331186-592a-4984-88de-70a2a27c9fb9).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.679990.

MLA Handbook (7^th Edition):

French, Joseph. “Towards an understanding of the burden of recombinant protein production.” 2016. Web. 24 Jan 2021.

Vancouver:

French J. Towards an understanding of the burden of recombinant protein production. [Internet] [Doctoral dissertation]. University of Manchester; 2016. [cited 2021 Jan 24]. Available from: https://www.research.manchester.ac.uk/portal/en/theses/towards-an-understanding-of-the-burden-of-recombinant-protein-production(4c331186-592a-4984-88de-70a2a27c9fb9).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.679990.

Council of Science Editors:

French J. Towards an understanding of the burden of recombinant protein production. [Doctoral Dissertation]. University of Manchester; 2016. Available from: https://www.research.manchester.ac.uk/portal/en/theses/towards-an-understanding-of-the-burden-of-recombinant-protein-production(4c331186-592a-4984-88de-70a2a27c9fb9).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.679990

University of Hawaii – Manoa

7. Kagawa, Allison. Towards the Development of a Recombinant Expression System for Yeast Reticulon Protein.

Degree: 2017, University of Hawaii – Manoa

URL: http://hdl.handle.net/10125/51169

►

M.S. University of Hawaii at Manoa 2015.

Structure determination by X-ray diffraction requires large amounts of purified, functional protein making it especially challenging for membrane… (more)

Subjects/Keywords: Reticulon; Recombinant; Membrane protein; Mass spectrometry

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APA (6^th Edition):

Kagawa, A. (2017). Towards the Development of a Recombinant Expression System for Yeast Reticulon Protein. (Thesis). University of Hawaii – Manoa. Retrieved from http://hdl.handle.net/10125/51169

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Kagawa, Allison. “Towards the Development of a Recombinant Expression System for Yeast Reticulon Protein.” 2017. Thesis, University of Hawaii – Manoa. Accessed January 24, 2021. http://hdl.handle.net/10125/51169.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Kagawa, Allison. “Towards the Development of a Recombinant Expression System for Yeast Reticulon Protein.” 2017. Web. 24 Jan 2021.

Vancouver:

Kagawa A. Towards the Development of a Recombinant Expression System for Yeast Reticulon Protein. [Internet] [Thesis]. University of Hawaii – Manoa; 2017. [cited 2021 Jan 24]. Available from: http://hdl.handle.net/10125/51169.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Kagawa A. Towards the Development of a Recombinant Expression System for Yeast Reticulon Protein. [Thesis]. University of Hawaii – Manoa; 2017. Available from: http://hdl.handle.net/10125/51169

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Universitat de Valencia

8. Martínez Solís, María. Interacción insecto-baculovirus. Aplicaciones en la mejora de baculovirus como vector para la expresión heteróloga de proteínas.

Degree: 2018, Universitat de Valencia

URL: http://hdl.handle.net/10550/66207

► Los baculovirus forman un amplio grupo de virus patógenos de insectos que se caracterizan por su elevada especificidad. Su principal aplicación es su uso como… (more)

▼ Los baculovirus forman un amplio grupo de virus patógenos de insectos que se caracterizan por su elevada especificidad. Su principal aplicación es su uso como agente bioinsecticida frente a diferentes plagas agrícolas, aunque algunas de sus características han permitido el desarrollo de tecnologías que permiten usar los baculovirus como vectores para la expresión heteróloga de proteínas, así como vectores para su uso en terapia génica. El sistema de expresión basado en baculovirus, conocido como BEVS (Baculovirus Expression Vector System), fue desarrollado en los años 80 y desde entonces ha permitido la expresión de gran diversidad de proteínas recombinantes en cultivos celulares de insectos, de una manera más rápida y económica que empleando otros sistemas de expresión. Sin embargo, este sistema de producción presenta una serie de limitaciones que afectan sobre todo cuando se quiere una producción a gran escala. Estas limitaciones suelen estar relacionadas con bajos rendimientos de expresión, disminuyendo así la eficiencia de producción. Además, la multiplicación continuada de los baculovirus en cultivos celulares da lugar a la aparición de virus defectivos que se caracterizan por la pérdida de partes de su genoma, pudiendo afectar a la producción de proteínas recombinantes por pérdida del gen recombinante correspondiente. Por ello, para que el sistema BEVS siga siendo un sistema de producción competitivo, es necesaria la introducción de mejoras que permitan aumentar los niveles de producción con respecto al sistema convencional. Aprovechando los conocimientos de estudios previos sobre la interacción de los baculovirus con su huésped, en esta tesis se ha abordado la introducción de mejoras en el sistema de expresión BEVS desde distintas perspectivas y desarrollando diferentes estrategias, que aparecen reflejadas en cada uno de los capítulos. En el primer capítulo se describe una secuencia derivada del genoma del baculovirus de S. exigua (SeMNPV) con actividad promotora, a partir de un gen viral altamente expresado en insectos infectados con dicho virus, a la que se ha denominado pSeL. Este nuevo promotor muestra unos niveles de expresión de la proteína GFP alrededor de 2 veces mayores con respecto a los obtenidos empleando el promotor del gen de la poliedrina (polh) en diferentes líneas celulares de insectos. Además, la combinación de los promotores pSeL y polh muestra un efecto aditivo frente a la expresión de ambos promotores empleados individualmente. En el segundo capítulo, el estudio transcripcional del proceso infectivo de los baculovirus AcMNPV y SeMNPV tanto en cultivos celulares como en larvas de S. exigua ha revelado un gen, el gen lef5, altamente sobreexpresado en larvas (respecto a su expresión en cultivo celular). Esto, nos dio pie a hipotetizar sobre su posible papel en la estabilidad genómica de los baculovirus durante su replicación en cultivo celular. El estudio de la sobreexpresión del gen lef5 de SeMNPV (Se-lef5) en baculovirus recombinantes, que además expresan GFP, confirma la influencia de… Advisors/Committee Members: Herrero Sendra, Salvador (advisor).

Subjects/Keywords: BEVS; baculovirus; expression system; recombinant protein

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Martínez Solís, M. (2018). Interacción insecto-baculovirus. Aplicaciones en la mejora de baculovirus como vector para la expresión heteróloga de proteínas. (Doctoral Dissertation). Universitat de Valencia. Retrieved from http://hdl.handle.net/10550/66207

Chicago Manual of Style (16^th Edition):

Martínez Solís, María. “Interacción insecto-baculovirus. Aplicaciones en la mejora de baculovirus como vector para la expresión heteróloga de proteínas. ” 2018. Doctoral Dissertation, Universitat de Valencia. Accessed January 24, 2021. http://hdl.handle.net/10550/66207.

MLA Handbook (7^th Edition):

Martínez Solís, María. “Interacción insecto-baculovirus. Aplicaciones en la mejora de baculovirus como vector para la expresión heteróloga de proteínas. ” 2018. Web. 24 Jan 2021.

Vancouver:

Martínez Solís M. Interacción insecto-baculovirus. Aplicaciones en la mejora de baculovirus como vector para la expresión heteróloga de proteínas. [Internet] [Doctoral dissertation]. Universitat de Valencia; 2018. [cited 2021 Jan 24]. Available from: http://hdl.handle.net/10550/66207.

Council of Science Editors:

Martínez Solís M. Interacción insecto-baculovirus. Aplicaciones en la mejora de baculovirus como vector para la expresión heteróloga de proteínas. [Doctoral Dissertation]. Universitat de Valencia; 2018. Available from: http://hdl.handle.net/10550/66207

University of Limerick

9. Witt, Madlen K. A study of selected environmental issues related o biopharmaceutical manufacturing using Escherichia coli to produce a recombinant protein.

Degree: 2014, University of Limerick

URL: http://hdl.handle.net/10344/4036

►

peer-reviewed

Escherichia coli expression systems remain a preferred choice for the production of recombinant proteins for therapeutic, diagnostic and industrial purposes. Low costs and simplicity… (more)

▼

peer-reviewed

Escherichia coli expression systems remain a preferred choice for the production of recombinant proteins for therapeutic, diagnostic and industrial purposes. Low costs and simplicity of culturing as well as straightforward genetic engineering technologies ensure their continued use for laboratory investigations as well as in commercial activities. An E. coli expression system producing a recombinant protein was constructed for this research. The model strain, E. coli MC106 producing recombinant bacterial His6-tagged β-galactosidase, was developed via standard genetic engineering techniques and protein expression was optimised to achieve high concentrations of soluble product. Historically, during upstream processing little consideration was given to the potential environmental impacts of culture media ingredients which were often added in excess to achieve high cell density and hence product yields. The model E. coli strain was utilised to investigate the scope for reducing phosphorus (P) quantities included in a complex (LB and TB) and semi-defined (M9/YE) fermentation media. The findings showed that P reductions of up to 70 % did not adversely affect biomass and product yields attained; however, further P minimisation lead to a drop in dry cell weight as well as protein synthesis, particularly in the case of semi-defined media. Protein functionality, assessed by the kinetic parameters Km and Vmax, was not influenced by the type of media nor the P concentration present. 70 % P reductions would lead to significant P savings in large-scale manufacturing of proteins produced by genetically engineered E. coli strains. The second part of this study entailed purification, at laboratory-scale to electrophoretic homogeneity, of the model protein via a traditional multichromatographic scheme and an affinity-based strategy. Both purification schemes were compared in terms of their environmental impact based on the buffers used. Utilising the engineered affinity-based approach reduced the number of downstream processing steps required to achieve purification from 6 to 4 and increased the final product yield from 11 % to 34 %. Environmental analysis of the chromatographic buffer constituents indicated that, per mg of purified protein, the use of the affinitybased method reduced the total P usage levels by 46 %, total ammonia by 99 %, total water usage by 75 % and total COD by 62 %, although the organic nitrogen levels increased by 75 %. In addition, comparative cost analysis showed a 60 % savings in chemical and chromatography costs per mg of purified product for this purification strategy. Although already widely used at research level, the use of affinity-based purification systems for process-scale protein purification would likely have significant environmental, energy and cost benefits. Furthermore, the study showed additional P savings can be achieved by using alternative buffering systems not containing P compounds during protein purification. Mass balance…

Advisors/Committee Members: Walsh, Gary, O'Dwyer, Tom, EPA.

Subjects/Keywords: Escherichia coli; recombinant protein; biotechnology industry

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APA (6^th Edition):

Witt, M. K. (2014). A study of selected environmental issues related o biopharmaceutical manufacturing using Escherichia coli to produce a recombinant protein. (Thesis). University of Limerick. Retrieved from http://hdl.handle.net/10344/4036

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Witt, Madlen K. “A study of selected environmental issues related o biopharmaceutical manufacturing using Escherichia coli to produce a recombinant protein.” 2014. Thesis, University of Limerick. Accessed January 24, 2021. http://hdl.handle.net/10344/4036.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Witt, Madlen K. “A study of selected environmental issues related o biopharmaceutical manufacturing using Escherichia coli to produce a recombinant protein.” 2014. Web. 24 Jan 2021.

Vancouver:

Witt MK. A study of selected environmental issues related o biopharmaceutical manufacturing using Escherichia coli to produce a recombinant protein. [Internet] [Thesis]. University of Limerick; 2014. [cited 2021 Jan 24]. Available from: http://hdl.handle.net/10344/4036.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Witt MK. A study of selected environmental issues related o biopharmaceutical manufacturing using Escherichia coli to produce a recombinant protein. [Thesis]. University of Limerick; 2014. Available from: http://hdl.handle.net/10344/4036

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Texas – Austin

10. Wallen, Michael Andrew, Jr. Isolation and characterization of Pisum sativum apyrases, PsNTP9 and PsNTP9-DM, cloned and expressed in Escherichia coli.

Degree: MA, Plant Biology, 2019, University of Texas – Austin

URL: http://dx.doi.org/10.26153/tsw/1436

► Adenosine triphosphate (ATP) is widely known as a fuel source for many biochemical processes, and to a lesser degree also as a signaling molecule in… (more)

▼ Adenosine triphosphate (ATP) is widely known as a fuel source for many biochemical processes, and to a lesser degree also as a signaling molecule in plants and animals. When plants are subjected to biotic or abiotic stress or undergoing exocytosis, they release ATP into the extracellular matrix (ECM). The release of ATP sets off a signal transduction pathway, first rapidly increasing the concentrations of cytosolic calcium, reactive oxygen species, and nitric oxide. How these changes specifically influence physiology is the object of much research in both plants and animals. Some of the changes that are affected influence growth and development, stomatal function, and gravitropism. Apyrases and other phosphatases control the concentration of the released nucleotides by breaking phosphate bonds from nucleoside triphosphates and diphosphates. Research aimed at the discovery of receptors, signaling pathway components, and processes has been successful to some extent. There are now known purinergic receptors in both plants and animal cells. We have cloned a truncated version of Pisum sativum (ps) NTP9. We used a pET-22B vector to add a histidine tag and transformed the vector into the BL21 Escherichia coli with a T7 promoter to enable IPTG induction of the LAC operon and expression of the enzyme. The pET-22B vector was incubated in separate samples with BL21 cells. Cells were propagated, and the expression of recombinant proteins PsNTP9, and separately, a double mutant PsNTP9-DM with a second calmodulin-binding domain, were induced ectopically. Cells were broken open by shaking them and mixing them with lysis buffer. Centrifugation was performed to separate the supernatant containing the released apyrases from the particulate wall fraction. The enzymes were purified by affinity chromatography, then their purity was evaluated by sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE). Western blots were performed to verify presence of the apyrases using a commercial anti-histidine antibody to detect PsNTP9 and PsNTP9-DM. Once suitable amounts of our proteins of interest were harvested, we performed Bradford assays to determine the protein concentration of the samples and carried out an apyrase activity assay to determine the specific activity of the purified enzymes and compare it to that of other known phosphatases. Advisors/Committee Members: Roux, Stanley J. (advisor).

Subjects/Keywords: Apyrase; eATP; Protein; Cloning; Recombinant; Fractionation; Chromatography

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APA (6^th Edition):

Wallen, Michael Andrew, J. (2019). Isolation and characterization of Pisum sativum apyrases, PsNTP9 and PsNTP9-DM, cloned and expressed in Escherichia coli. (Masters Thesis). University of Texas – Austin. Retrieved from http://dx.doi.org/10.26153/tsw/1436

Chicago Manual of Style (16^th Edition):

Wallen, Michael Andrew, Jr. “Isolation and characterization of Pisum sativum apyrases, PsNTP9 and PsNTP9-DM, cloned and expressed in Escherichia coli.” 2019. Masters Thesis, University of Texas – Austin. Accessed January 24, 2021. http://dx.doi.org/10.26153/tsw/1436.

MLA Handbook (7^th Edition):

Wallen, Michael Andrew, Jr. “Isolation and characterization of Pisum sativum apyrases, PsNTP9 and PsNTP9-DM, cloned and expressed in Escherichia coli.” 2019. Web. 24 Jan 2021.

Vancouver:

Wallen, Michael Andrew J. Isolation and characterization of Pisum sativum apyrases, PsNTP9 and PsNTP9-DM, cloned and expressed in Escherichia coli. [Internet] [Masters thesis]. University of Texas – Austin; 2019. [cited 2021 Jan 24]. Available from: http://dx.doi.org/10.26153/tsw/1436.

Council of Science Editors:

Wallen, Michael Andrew J. Isolation and characterization of Pisum sativum apyrases, PsNTP9 and PsNTP9-DM, cloned and expressed in Escherichia coli. [Masters Thesis]. University of Texas – Austin; 2019. Available from: http://dx.doi.org/10.26153/tsw/1436

Purdue University

11. Kim, Yeji. MODULATING CELL DIFFERENTIATION WITH PROTEIN-ENGINEERED MICROENVIRONMENTS.

Degree: PhD, Chemical Engineering, 2014, Purdue University

URL: https://docs.lib.purdue.edu/open_access_dissertations/1070

► Tissue damage caused by diseases and injuries requires regeneration to recover proper tissue function. Traditional treatments using autologous tissue have not yet met the need… (more)

Subjects/Keywords: Biomaterials; Microenvironments; Recombinant protein; Stem cell differentiation

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APA (6^th Edition):

Kim, Y. (2014). MODULATING CELL DIFFERENTIATION WITH PROTEIN-ENGINEERED MICROENVIRONMENTS. (Doctoral Dissertation). Purdue University. Retrieved from https://docs.lib.purdue.edu/open_access_dissertations/1070

Chicago Manual of Style (16^th Edition):

Kim, Yeji. “MODULATING CELL DIFFERENTIATION WITH PROTEIN-ENGINEERED MICROENVIRONMENTS.” 2014. Doctoral Dissertation, Purdue University. Accessed January 24, 2021. https://docs.lib.purdue.edu/open_access_dissertations/1070.

MLA Handbook (7^th Edition):

Kim, Yeji. “MODULATING CELL DIFFERENTIATION WITH PROTEIN-ENGINEERED MICROENVIRONMENTS.” 2014. Web. 24 Jan 2021.

Vancouver:

Kim Y. MODULATING CELL DIFFERENTIATION WITH PROTEIN-ENGINEERED MICROENVIRONMENTS. [Internet] [Doctoral dissertation]. Purdue University; 2014. [cited 2021 Jan 24]. Available from: https://docs.lib.purdue.edu/open_access_dissertations/1070.

Council of Science Editors:

Kim Y. MODULATING CELL DIFFERENTIATION WITH PROTEIN-ENGINEERED MICROENVIRONMENTS. [Doctoral Dissertation]. Purdue University; 2014. Available from: https://docs.lib.purdue.edu/open_access_dissertations/1070

San Jose State University

12. Chen, Melvin. Effects of mlc Gene Modulation on Acetate Accumulation in Escherichia Coli Culture.

Degree: MS, Biomedical, Chemical & Materials Engineering, 2014, San Jose State University

URL: https://doi.org/10.31979/etd.2qv8-qcbe ; https://scholarworks.sjsu.edu/etd_theses/4491

► When Escherichia coli (E. coli) is grown in the presence of excess glucose, acetate is produced, oftentimes as an undesired by-product. Mlc is a… (more)

Subjects/Keywords: E. Coli; glucose; mlc; modulation; recombinant protein; therapeutic protein

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APA (6^th Edition):

Chen, M. (2014). Effects of mlc Gene Modulation on Acetate Accumulation in Escherichia Coli Culture. (Masters Thesis). San Jose State University. Retrieved from https://doi.org/10.31979/etd.2qv8-qcbe ; https://scholarworks.sjsu.edu/etd_theses/4491

Chicago Manual of Style (16^th Edition):

Chen, Melvin. “Effects of mlc Gene Modulation on Acetate Accumulation in Escherichia Coli Culture.” 2014. Masters Thesis, San Jose State University. Accessed January 24, 2021. https://doi.org/10.31979/etd.2qv8-qcbe ; https://scholarworks.sjsu.edu/etd_theses/4491.

MLA Handbook (7^th Edition):

Chen, Melvin. “Effects of mlc Gene Modulation on Acetate Accumulation in Escherichia Coli Culture.” 2014. Web. 24 Jan 2021.

Vancouver:

Chen M. Effects of mlc Gene Modulation on Acetate Accumulation in Escherichia Coli Culture. [Internet] [Masters thesis]. San Jose State University; 2014. [cited 2021 Jan 24]. Available from: https://doi.org/10.31979/etd.2qv8-qcbe ; https://scholarworks.sjsu.edu/etd_theses/4491.

Council of Science Editors:

Chen M. Effects of mlc Gene Modulation on Acetate Accumulation in Escherichia Coli Culture. [Masters Thesis]. San Jose State University; 2014. Available from: https://doi.org/10.31979/etd.2qv8-qcbe ; https://scholarworks.sjsu.edu/etd_theses/4491

University of Western Ontario

13. Lama, Pravesh. Identification and Functional Characterization of GmMYB176-Specific Protein Kinases in Soybean.

Degree: 2016, University of Western Ontario

URL: https://ir.lib.uwo.ca/etd/3823

► GmMYB176 is an R1 MYB transcription factor that regulates isoflavonoid biosynthesis in soybean. The deletion of phosphorylation sites in GmMYB176 abrogates its interaction with 14-3-3… (more)

Subjects/Keywords: Isoflavonoid; protein kinase; recombinant protein expression; kinase assay; Biology; Life Sciences

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APA (6^th Edition):

Lama, P. (2016). Identification and Functional Characterization of GmMYB176-Specific Protein Kinases in Soybean. (Thesis). University of Western Ontario. Retrieved from https://ir.lib.uwo.ca/etd/3823

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Lama, Pravesh. “Identification and Functional Characterization of GmMYB176-Specific Protein Kinases in Soybean.” 2016. Thesis, University of Western Ontario. Accessed January 24, 2021. https://ir.lib.uwo.ca/etd/3823.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Lama, Pravesh. “Identification and Functional Characterization of GmMYB176-Specific Protein Kinases in Soybean.” 2016. Web. 24 Jan 2021.

Vancouver:

Lama P. Identification and Functional Characterization of GmMYB176-Specific Protein Kinases in Soybean. [Internet] [Thesis]. University of Western Ontario; 2016. [cited 2021 Jan 24]. Available from: https://ir.lib.uwo.ca/etd/3823.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Lama P. Identification and Functional Characterization of GmMYB176-Specific Protein Kinases in Soybean. [Thesis]. University of Western Ontario; 2016. Available from: https://ir.lib.uwo.ca/etd/3823

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Universitat Autònoma de Barcelona

14. Adelantado Vallvé, Núria. Lipidomics studies of recombinant Pichia pastoris for improved recombinant protein secretion through cell engineering.

Degree: Departament d'Enginyeria Química, 2016, Universitat Autònoma de Barcelona

URL: http://hdl.handle.net/10803/384229

► The increasing availability of omics databases provides an important knowledge base for the design of novel yeast engineering strategies, since they offer systems-level information on… (more)

▼ The increasing availability of omics databases provides an important knowledge base for the design of novel yeast engineering strategies, since they offer systems-level information on the physiology of the cells under diverse growth conditions and genetic backgrounds. In a previous study, a beneficial effect of low oxygen availability on the expression of a human Fab fragment in Pichia pastoris has been identified. Transcriptomic analyses pointed out important regulation of the Unfolded Protein Response (UPR) and lipid metabolism, with a significant transcriptional up-regulation of ergosterol and sphingolipid biosynthetic pathways. Furthermore, cells cultured in the presence of fluconazole, an antifungal agent that inhibits sterol C14α-demethylase (Erg11p) activity in the ergosterol biosynthetic pathway, showed a reduced amount of ergosterol in the plasma membrane, resulting in increased protein secretion levels. Moreover, it is known that the transcription factor Hac1, which plays a key role by regulating the UPR, regulates lipid biosynthesis In the present study, a reference strain of P. pastoris secreting the 2F5 Fab antibody fragment as a model recombinant protein was genetically modified in order to change its lipid composition. A set of strains harbouring either deleted (DES1, SUR2) or overexpressed (DES1) genes of the sphingolipid pathway were generated. In addition, a Fab producing strain overexpressing HAC1 or ERG11 were constructed and, as an alternative to the ergosterol pathway knockout, the reference strain was treated with fluconazole. The series of strains were cultivated in chemostat cultures under normoxic and hypoxic conditions. Strains were characterised in terms of Fab- productivity, intra/extracellular product distribution, cell morphology by transmission electron microscopy (TEM) and lipid content in terms of phospholipids, fatty acids, sterols, non-polar lipids and sphingolipids, in order to fully understand how cells adapt to genetic and environmental changes involving lipid changes. The obtained results were combined with previously published transcriptional data, allowing us to demonstrate an important remodelling of the lipid metabolism under limited oxygen availability. Advisors/Committee Members: [email protected] (authoremail), true (authoremailshow), Ferrer, Pau (director), Valero, Francisco (director), true (authorsendemail).

Subjects/Keywords: Lípid; Lípido; Lipid; Pichia Pastoris; Proteïna recombinant; Proteína recominante; Recombinant protein; Tecnologies; 57

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Adelantado Vallvé, N. (2016). Lipidomics studies of recombinant Pichia pastoris for improved recombinant protein secretion through cell engineering. (Thesis). Universitat Autònoma de Barcelona. Retrieved from http://hdl.handle.net/10803/384229

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Adelantado Vallvé, Núria. “Lipidomics studies of recombinant Pichia pastoris for improved recombinant protein secretion through cell engineering.” 2016. Thesis, Universitat Autònoma de Barcelona. Accessed January 24, 2021. http://hdl.handle.net/10803/384229.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Adelantado Vallvé, Núria. “Lipidomics studies of recombinant Pichia pastoris for improved recombinant protein secretion through cell engineering.” 2016. Web. 24 Jan 2021.

Vancouver:

Adelantado Vallvé N. Lipidomics studies of recombinant Pichia pastoris for improved recombinant protein secretion through cell engineering. [Internet] [Thesis]. Universitat Autònoma de Barcelona; 2016. [cited 2021 Jan 24]. Available from: http://hdl.handle.net/10803/384229.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Adelantado Vallvé N. Lipidomics studies of recombinant Pichia pastoris for improved recombinant protein secretion through cell engineering. [Thesis]. Universitat Autònoma de Barcelona; 2016. Available from: http://hdl.handle.net/10803/384229

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

15. Michelle Rossana Ferreira Vaz. Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi.

Degree: 2008, Universidade Federal do Rio Grande do Norte

URL: http://bdtd.bczm.ufrn.br/tedesimplificado//tde_busca/arquivo.php?codArquivo=1662

►

Com advento da tecnologia do DNA recombinante, a expressão de proteínas recombinantes torna-se uma ferramenta importante nos estudos da estrutura, função e identificação de novas… (more)

▼

Com advento da tecnologia do DNA recombinante, a expressão de proteínas recombinantes torna-se uma ferramenta importante nos estudos da estrutura, função e identificação de novas proteínas, principalmente com finalidades terapêuticas. A Escherichia coli tem sido o procarioto predominante nos estudos da engenharia genética devido à riqueza de informações a respeito do seu metabolismo. Apesar do avanço expressivo dos estudos da biologia molecular e da imunologia das infecções, não existe, atualmente, nenhuma droga profilática capaz de prevenir o calazar. Desta forma, existe uma grande necessidade de identificação de antígenos específicos para o desenvolvimento de vacinas e kits para diagnósticos contra a Leishmaniose visceral. Neste contexto, este trabalho objetivou estudar a expressão de antígenos recombinantes da Leishmania chagasi durante o cultivo de Escherichia coli em incubador rotativo (shaker). Um primeiro conjunto de ensaios foi realizado com o objetivo de se conhecer o comportamento cinético do crescimento dois clones recombinantes (eIF, LACK) em duas diferentes composições de meios (2xTY, TB) suplementados por antibióticos, sem adição de IPTG. Na segunda etapa dos ensaios, foi realizado o procedimento de indução por IPTG, a fim de verificar a influência da composição dos meios testados na expressão das proteínas recombinantes. Com base nos resultados obtidos, pode-se observar que a elevada complexidade do meio de cultivo favoreceu a cinética de crescimento dos clones recombinantes (eIF,LACK), no entanto, ao se tratar dos ensaios submetidos ao procedimento de indução por IPTG, a elevada complexidade do meio de cultivo não favoreceu a expressão das proteínas recombinantes. Por outro lado, foram obtidos resultados positivos para todos os clones recombinante (eIF, LACK) testados, confirmada através do perfil eletroforético

With advent of the technology of the recombinant DNA, the recombinant protein expression becomes an important tool in the studies of the structure, function and identification of new proteins, mainly with therapeutical purposes. The Escherichia coli has been procarioto predominant in the studies of genetic engineering due to wealth of information regarding its metabolism. Despite the expressivo advance of the studies of molecular biology and the immunology of the infections, it does not exist, currently, no prophylactic drug capable to prevent calazar. Of this form, it exists a great necessity of specific antigen identification for the vaccine development and kits for disgnostic against the visceral Leishmaniose. In this context, this work objectified to study the recombinant antigen expression of the Leishmania chagasi during the culture of Escherichia coli in shaker. A first set of assays was carried through with the objective of if knowing the kinetic behavior of the growth of two clones recombinant proteins (eIF, LACK) in two different compositions of culture medium (2xTY, TB) supplemented by antibiotics, without IPTG addition. In the second stage of the assays, the procedure of…

Advisors/Committee Members: Márcia Regina da Silva Pedrini, Everaldo Silvino dos Santos, Gorete Ribeiro de Macedo, Ester Ribeiro Gouveia.

Subjects/Keywords: Proteína; Recombinante; Leishmaniose visceral; ENGENHARIA QUIMICA; Protein; Recombinant; Visceral leishmaniasis

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Vaz, M. R. F. (2008). Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi. (Thesis). Universidade Federal do Rio Grande do Norte. Retrieved from http://bdtd.bczm.ufrn.br/tedesimplificado//tde_busca/arquivo.php?codArquivo=1662

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Vaz, Michelle Rossana Ferreira. “Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi.” 2008. Thesis, Universidade Federal do Rio Grande do Norte. Accessed January 24, 2021. http://bdtd.bczm.ufrn.br/tedesimplificado//tde_busca/arquivo.php?codArquivo=1662.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Vaz, Michelle Rossana Ferreira. “Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi.” 2008. Web. 24 Jan 2021.

Vancouver:

Vaz MRF. Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi. [Internet] [Thesis]. Universidade Federal do Rio Grande do Norte; 2008. [cited 2021 Jan 24]. Available from: http://bdtd.bczm.ufrn.br/tedesimplificado//tde_busca/arquivo.php?codArquivo=1662.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Vaz MRF. Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi. [Thesis]. Universidade Federal do Rio Grande do Norte; 2008. Available from: http://bdtd.bczm.ufrn.br/tedesimplificado//tde_busca/arquivo.php?codArquivo=1662

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of California – Berkeley

16. Dana, Craig Matthew. Cel7A Engineering and Expression.

Degree: Chemical Engineering, 2013, University of California – Berkeley

URL: http://www.escholarship.org/uc/item/3sb6n3n5

► Renewable fuels produced from biomass-derived sugars are receiving increasing attention. Lignocellulose-degrading enzymes derived from fungi are attractive for saccharification of biomass because they can be… (more)

▼ Renewable fuels produced from biomass-derived sugars are receiving increasing attention. Lignocellulose-degrading enzymes derived from fungi are attractive for saccharification of biomass because they can be produced at higher titers and at significantly less cost than those produced by bacteria or archaea. However, their properties can be suboptimal; for example, they are subject to product inhibition and are sensitive to small changes in pH. Furthermore, increased thermostability would be advantageous for saccharification as increased temperature may reduce the risk of microbial contamination. Therefore, there is a need for a generalized platform that can be applied to the engineering of these enzymes. Commercially available lignocellulose-degrading enzymes are produced using a hypersecreting strain of the filamentous fungus Trichoderma reesei. Among the enzymes secreted by this organism, the cellulase Cel7A is present in the highest concentration and is the only enzyme responsible for non-reducing end directed exo-acting cellulolytic activity. Additionally, the enzyme's presence is critical for growth of T. reesei on cellulosic substrates. Here, a general mutagenesis platform that employed the budding yeast Saccharomyces cerevisiae was developed to improve the properties of Cel7A. Secretion of Cel7A at titers of 26 mg/L with limited hyperglycosylation was achieved using an S. cerevisiae strain with upregulated protein disulfide isomerase, an engineered α-factor prepro leader, and the deletion of a plasma membrane ATPase. Because cellulase activities are difficult to screen in high-throughput, a DNA shuffling based library generation technique that results in a high percentage of active clones was developed called Biased Clique Shuffling (BCS). BCS allows for the control of DNA diversity during library generation. Applying this technique to 11 homologous Cel7A genes, we generated several libraries that were rich in activity and identified chimeras with improved thermostability, thermal activity, and product inhibition. The libraries generated using the BCS technique were far superior as a source of active and stable chimeras compared to an equimolar library prepared from the same 11 genes (as is classically prepared using DNA shuffling). Finally, we found that Cel7A expressed in the filamentous fungus N. crassa had twice the specific activity at 65°C and a 10°C higher Tm relative to Cel7A expressed in S. cerevisiae. Through a study of the three known post-translational modifications, namely glycosylation, disulfide bond formation, and N-terminal glutamine cyclization, we revealed that S. cerevisiae expressed Cel7A with an unmodified N-terminus, unlike native Cel7A which has an N-terminal pyroglutamate. Furthermore cyclizing the unmodified N-terminal glutamine in the Cel7A expressed in S. cerevisiae to form pryoglutamate in-vitro with glutaminyl cyclase increased the enzyme's specific activity and thermostability to match those Cel7A expressed in N. crassa. This unprecedented result…

Subjects/Keywords: Chemical engineering; CBHI; Cel7A; cellulase; protein engineering; recombinant expression; saccharomyces cerevisiae

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Dana, C. M. (2013). Cel7A Engineering and Expression. (Thesis). University of California – Berkeley. Retrieved from http://www.escholarship.org/uc/item/3sb6n3n5

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Dana, Craig Matthew. “Cel7A Engineering and Expression.” 2013. Thesis, University of California – Berkeley. Accessed January 24, 2021. http://www.escholarship.org/uc/item/3sb6n3n5.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Dana, Craig Matthew. “Cel7A Engineering and Expression.” 2013. Web. 24 Jan 2021.

Vancouver:

Dana CM. Cel7A Engineering and Expression. [Internet] [Thesis]. University of California – Berkeley; 2013. [cited 2021 Jan 24]. Available from: http://www.escholarship.org/uc/item/3sb6n3n5.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Dana CM. Cel7A Engineering and Expression. [Thesis]. University of California – Berkeley; 2013. Available from: http://www.escholarship.org/uc/item/3sb6n3n5

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

17. Comenale, Gabriela. Expressão e purificação da proteína recombinante L2 do Papilomavírus bovino tipo-2 em sistema bacteriano.

Degree: Mestrado, Anatomia dos Animais Domésticos e Silvestres, 2012, University of São Paulo

URL: http://www.teses.usp.br/teses/disponiveis/10/10132/tde-11102013-104819/ ;

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A papilomatose bovina é uma doença infectocontagiosa de ocorrência mundial, que assola o rebanho brasileiro, sem qualquer atitude efetiva de controle, e que tem como… (more)

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A papilomatose bovina é uma doença infectocontagiosa de ocorrência mundial, que assola o rebanho brasileiro, sem qualquer atitude efetiva de controle, e que tem como enfermidades associadas a tumores de bexiga hematúria enzoótica e tumores de trato digestório superior caraguatá, responsáveis por sensíveis perdas para a pecuária. Várias tentativas vacinais têm sido empreendidas com finalidades profiláticas ou terapêuticas, porém sem resultados eficazes. Esta situação se deve a aspectos relacionados à estrutura viral que dificultam uma manipulação eficiente para produção de produtos vacinais. Para que tais dados possam ser obtidos, faz-se necessário uma melhor compreensão da ação das proteínas recombinantes. A clonagem em vetores bacterianos para a expressão e purificação dessas proteínas serve a diferentes propósitos. Entre eles, a produção de insumos imunológicos, como testes diagnósticos ou mesmo vacinas. O presente projeto visou à expressão e purificação da proteína de capsídeo recombinante L2 de BPV-2. A proteína foi expressa em bactéria e tentou-se a purificação por coluna de afinidade; entretanto, problemas na purificação não possibilitaram a conclusão deste objetivo. Todas as abordagens e protocolos utilizados nesse trabalho são discutidos para contribuição ao conhecimento do processo.

The bovine papillomatosis is an infectious disease of worldwide occurrence, plaguing the Brazilian herd, without any effective attitude control, and whose illnesses associated with bladder tumors ënzootic hematuriaänd upper digestive tract tumors \"caraguatás̈ensitive responsible for losses to livestock. Several attempts have been undertaken vaccine with prophylactic or therapeutic purposes, but without effective results. This is due to issues related to viral structure that hinder efficient manipulation for production of vaccine products. In order to obtain such information, it is necessary better understanding of the action of recombinant proteins. The bacterial cloning vectors for the expression and purification of such proteins serve different purposes. Among them, the production of immune inputs, such as diagnostic tests or vaccines. This project aimed the expression and purification of recombinant L2 capsid protein of BPV-2. The protein was expressed in bacteria and purification was carried out by affinity column. However, difficulties in the purification process, impaired the full completion of this objective. All the attempted approaches and protocols were discussed and potential solutions proposed.

Advisors/Committee Members: Kfoury Junior, José Roberto.

Subjects/Keywords: Bovine papilomavírus; Papilomavírus bovino; Proteína recombinante; Recombinant protein

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Comenale, G. (2012). Expressão e purificação da proteína recombinante L2 do Papilomavírus bovino tipo-2 em sistema bacteriano. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/10/10132/tde-11102013-104819/ ;

Chicago Manual of Style (16^th Edition):

Comenale, Gabriela. “Expressão e purificação da proteína recombinante L2 do Papilomavírus bovino tipo-2 em sistema bacteriano.” 2012. Masters Thesis, University of São Paulo. Accessed January 24, 2021. http://www.teses.usp.br/teses/disponiveis/10/10132/tde-11102013-104819/ ;.

MLA Handbook (7^th Edition):

Comenale, Gabriela. “Expressão e purificação da proteína recombinante L2 do Papilomavírus bovino tipo-2 em sistema bacteriano.” 2012. Web. 24 Jan 2021.

Vancouver:

Comenale G. Expressão e purificação da proteína recombinante L2 do Papilomavírus bovino tipo-2 em sistema bacteriano. [Internet] [Masters thesis]. University of São Paulo; 2012. [cited 2021 Jan 24]. Available from: http://www.teses.usp.br/teses/disponiveis/10/10132/tde-11102013-104819/ ;.

Council of Science Editors:

Comenale G. Expressão e purificação da proteína recombinante L2 do Papilomavírus bovino tipo-2 em sistema bacteriano. [Masters Thesis]. University of São Paulo; 2012. Available from: http://www.teses.usp.br/teses/disponiveis/10/10132/tde-11102013-104819/ ;

18. Corgozinho, Carolina Nunes Costa. Desenvolvimento de vacina baseada em sistema de liberação sustentada contendo proteína recombinante.

Degree: Mestrado, Medicamentos e Cosméticos, 2008, University of São Paulo

URL: http://www.teses.usp.br/teses/disponiveis/60/60137/tde-31072009-083709/ ;

►

No Brasil, e em outros paises de clima tropical, os carrapatos se tornaram um enorme problema economico de^de que a industria do gado se desenvolveu.… (more)

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No Brasil, e em outros paises de clima tropical, os carrapatos se tornaram um enorme problema economico de^de que a industria do gado se desenvolveu. O carrapato Boophilus microplus, um dos artropodes mais importantes na veterinaria, causa efeitos direto, como suc,cao de sangue, e indireto, como a transmissao de uma grande variedade de patogenos que normalmente resulta em infec,c~es letais. As vacinas genicas contendo o antigeno Bm86, uma proteina ligada a membrana do intestino do carrapato B. microplus, representam uma alternativa atrativa aos acaricidas para controlar as infesta~oes por carrapatos em contrapartida aos inconvenientes produtos quimicos. Devido sua administra,cao ser feita em 4 doses no primeiro ano, seguida de refor,cos a cada seis meses, estas formula,coes vacinais nao s3c adequadas para paises com cria,cao extensiva de gado, como no Brasil. Visando uma libera~ao sustentada do antigeno Bm86, neste trabalho desenvolveuse uma vacina de dose unica baseada em microesferas polimericas. Para obter o padrao de liberac,ao desejavel, diferentes formula,coes e parametros de processo foram variados, como a composi,cao do polimero, a taxa entre os monomeros ^Uacido latico:acido glicolicoë o tamanho das microparticulas. As formula,coes foram preparadas pelo metodo de emulsao multipla e evapora,cao do solvente. A formula~ao que melhor se enquadrou nos objetivos da vacina de dose unica foi preparada com PLGA 75:25, solu,cao 3% de PVA como estabilizante, agita,cao de 11000 rpm para forma,cao da emulsao primaria e de 800 rpm para forma,cao da emulsao multipla e evapora,cao do solvente. As particulas assim obtidas apresentaram um tamanho medio de 25 ,um, uma taxa de encapsula,cao maior que 90% e aproximadamente 50% da proteina foi liberada in vitro em 60 dias. Analises por SDS-PAGE e Westem Bloning revelaram que a proteina se manteve integra apos encapsula,cao. Os resultados da avalia,cao da imunogenicidade em bovinos mostraram que a formula,cao baseada em microesferas polimericas biodegradaveis e habil a conseguir, com uma unica dose, uma resposta imune similar aquela conseguida com tres doses das formula,coes convencionais da vacina de Bm86.

In Brazil, and in others tropical countries, the ticks have become a huge economic problem since the industry of livestock has developed. Ticks and tick-borne diseases affect animal and human health and are the cause of significant economic losses. The cattle tick Boophilus microplus is one of the most important arthropods in veterinary. This tick species causes both direct effects, such as blood sucking, and indirect effects, such as transmission of a wide variety of pathogens, which usually result in lethal infections. The gene vaccines based on Bm86 antigen, a midgut membrane-bound protein of the cattle tick B. microplus, represent a good alternative to control tick infestations, compared to chemicals. However, due to these vaccine formulations need 4 doses over the first year with booster at each 6 months to be effective, they are not suitable for countries with…

Advisors/Committee Members: Bentley, Maria Vitoria Lopes Badra.

Subjects/Keywords: Bm86; Bm86; Microesfera; Microspheres; Proteina recombinante; Recombinant protein; Vaccine; Vacina

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Corgozinho, C. N. C. (2008). Desenvolvimento de vacina baseada em sistema de liberação sustentada contendo proteína recombinante. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/60/60137/tde-31072009-083709/ ;

Chicago Manual of Style (16^th Edition):

Corgozinho, Carolina Nunes Costa. “Desenvolvimento de vacina baseada em sistema de liberação sustentada contendo proteína recombinante.” 2008. Masters Thesis, University of São Paulo. Accessed January 24, 2021. http://www.teses.usp.br/teses/disponiveis/60/60137/tde-31072009-083709/ ;.

MLA Handbook (7^th Edition):

Corgozinho, Carolina Nunes Costa. “Desenvolvimento de vacina baseada em sistema de liberação sustentada contendo proteína recombinante.” 2008. Web. 24 Jan 2021.

Vancouver:

Corgozinho CNC. Desenvolvimento de vacina baseada em sistema de liberação sustentada contendo proteína recombinante. [Internet] [Masters thesis]. University of São Paulo; 2008. [cited 2021 Jan 24]. Available from: http://www.teses.usp.br/teses/disponiveis/60/60137/tde-31072009-083709/ ;.

Council of Science Editors:

Corgozinho CNC. Desenvolvimento de vacina baseada em sistema de liberação sustentada contendo proteína recombinante. [Masters Thesis]. University of São Paulo; 2008. Available from: http://www.teses.usp.br/teses/disponiveis/60/60137/tde-31072009-083709/ ;

Universidade do Rio Grande do Norte

19. Vaz, Michelle Rossana Ferreira. Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi .

Degree: 2008, Universidade do Rio Grande do Norte

URL: http://repositorio.ufrn.br/handle/123456789/15741

► With advent of the technology of the recombinant DNA, the recombinant protein expression becomes an important tool in the studies of the structure, function and… (more)

▼ With advent of the technology of the recombinant DNA, the recombinant protein expression becomes an important tool in the studies of the structure, function and identification of new proteins, mainly with therapeutical purposes. The Escherichia coli has been procarioto predominant in the studies of genetic engineering due to wealth of information regarding its metabolism. Despite the expressivo advance of the studies of molecular biology and the immunology of the infections, it does not exist, currently, no prophylactic drug capable to prevent calazar. Of this form, it exists a great necessity of specific antigen identification for the vaccine development and kits for disgnostic against the visceral Leishmaniose. In this context, this work objectified to study the recombinant antigen expression of the Leishmania chagasi during the culture of Escherichia coli in shaker. A first set of assays was carried through with the objective of if knowing the kinetic behavior of the growth of two clones recombinant proteins (eIF, LACK) in two different compositions of culture medium (2xTY, TB) supplemented by antibiotics, without IPTG addition. In the second stage of the assays, the procedure of induction for IPTG was carried through, in order to verify the influence of the composition of the ways tested in the expression them recombinant proteins. On the basis of the gotten results, can be observed that the high complexity of culture medium favored the kinetic one of growth of clones recombinant (eIF, LACK), however, to if to deal with the assays submitted to the procedure of induction for IPTG, the raised complexity of culture medium did not favor the expression of recombinant proteins. On the other hand, they had been gotten resulted positive for all clones recombinant (eIF, LACK) tested, confirmed through the eletroforético profile Advisors/Committee Members: Macedo, Gorete Ribeiro de (advisor), CPF:10847022404 (advisor), http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788057U7 (advisor), Pedrini, Márcia Regina da Silva (advisor), CPF:85203513953 (advisor), http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797345U9 (advisor).

Subjects/Keywords: Proteína; Recombinante; Leishmaniose visceral; Protein; Recombinant; Visceral leishmaniasis

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❌

APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Vaz, M. R. F. (2008). Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi . (Masters Thesis). Universidade do Rio Grande do Norte. Retrieved from http://repositorio.ufrn.br/handle/123456789/15741

Chicago Manual of Style (16^th Edition):

Vaz, Michelle Rossana Ferreira. “Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi .” 2008. Masters Thesis, Universidade do Rio Grande do Norte. Accessed January 24, 2021. http://repositorio.ufrn.br/handle/123456789/15741.

MLA Handbook (7^th Edition):

Vaz, Michelle Rossana Ferreira. “Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi .” 2008. Web. 24 Jan 2021.

Vancouver:

Vaz MRF. Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi . [Internet] [Masters thesis]. Universidade do Rio Grande do Norte; 2008. [cited 2021 Jan 24]. Available from: http://repositorio.ufrn.br/handle/123456789/15741.

Council of Science Editors:

Vaz MRF. Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi . [Masters Thesis]. Universidade do Rio Grande do Norte; 2008. Available from: http://repositorio.ufrn.br/handle/123456789/15741

Universidade do Rio Grande do Norte

20. Vaz, Michelle Rossana Ferreira. Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi .

Degree: 2008, Universidade do Rio Grande do Norte

URL: http://repositorio.ufrn.br/handle/123456789/15741

► With advent of the technology of the recombinant DNA, the recombinant protein expression becomes an important tool in the studies of the structure, function and… (more)

▼ With advent of the technology of the recombinant DNA, the recombinant protein expression becomes an important tool in the studies of the structure, function and identification of new proteins, mainly with therapeutical purposes. The Escherichia coli has been procarioto predominant in the studies of genetic engineering due to wealth of information regarding its metabolism. Despite the expressivo advance of the studies of molecular biology and the immunology of the infections, it does not exist, currently, no prophylactic drug capable to prevent calazar. Of this form, it exists a great necessity of specific antigen identification for the vaccine development and kits for disgnostic against the visceral Leishmaniose. In this context, this work objectified to study the recombinant antigen expression of the Leishmania chagasi during the culture of Escherichia coli in shaker. A first set of assays was carried through with the objective of if knowing the kinetic behavior of the growth of two clones recombinant proteins (eIF, LACK) in two different compositions of culture medium (2xTY, TB) supplemented by antibiotics, without IPTG addition. In the second stage of the assays, the procedure of induction for IPTG was carried through, in order to verify the influence of the composition of the ways tested in the expression them recombinant proteins. On the basis of the gotten results, can be observed that the high complexity of culture medium favored the kinetic one of growth of clones recombinant (eIF, LACK), however, to if to deal with the assays submitted to the procedure of induction for IPTG, the raised complexity of culture medium did not favor the expression of recombinant proteins. On the other hand, they had been gotten resulted positive for all clones recombinant (eIF, LACK) tested, confirmed through the eletroforético profile Advisors/Committee Members: Macedo, Gorete Ribeiro de (advisor), CPF:10847022404 (advisor), http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788057U7 (advisor), Pedrini, Márcia Regina da Silva (advisor), CPF:85203513953 (advisor), http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797345U9 (advisor).

Subjects/Keywords: Proteína; Recombinante; Leishmaniose visceral; Protein; Recombinant; Visceral leishmaniasis

Record Details Similar Records Cite « Share

❌

APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Vaz, M. R. F. (2008). Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi . (Thesis). Universidade do Rio Grande do Norte. Retrieved from http://repositorio.ufrn.br/handle/123456789/15741

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Vaz, Michelle Rossana Ferreira. “Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi .” 2008. Thesis, Universidade do Rio Grande do Norte. Accessed January 24, 2021. http://repositorio.ufrn.br/handle/123456789/15741.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Vaz, Michelle Rossana Ferreira. “Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi .” 2008. Web. 24 Jan 2021.

Vancouver:

Vaz MRF. Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi . [Internet] [Thesis]. Universidade do Rio Grande do Norte; 2008. [cited 2021 Jan 24]. Available from: http://repositorio.ufrn.br/handle/123456789/15741.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Vaz MRF. Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi . [Thesis]. Universidade do Rio Grande do Norte; 2008. Available from: http://repositorio.ufrn.br/handle/123456789/15741

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Waterloo

21. Yaeck, Jason. Production and Modeling of Recombinant Protein.

Degree: 2007, University of Waterloo

URL: http://hdl.handle.net/10012/2661

► Computational models of recombinant production of tissue-type Plasminogen Activator (tPA) were created, studied and compared for two hosts, Chinese Hamster Ovary (CHO)cells and Escherichia coli… (more)

▼ Computational models of recombinant production of tissue-type Plasminogen Activator (tPA) were created, studied and compared for two hosts, Chinese Hamster Ovary (CHO)cells and Escherichia coli (E. coli), using SuperPro® Designer. In addition, several fermentations were run using enhanced Yellow Fluorescent Protein (eYFP) in E. coli to provide knowledge for the SuperPro model and to explore the effect of temperature when used to maintain dissolved oxygen in a high density fed-batch fermentation. The models show that production of tPA is feasible using either host, but under the current basecase CHO holds the economic advantage despite the initial higher capital costs. In order to become more competitive with CHO, production using E. coli must become higher on a cell specific level and the potential of refolding insoluble protein in inclusion bodies should be explored. Since E. coli’s growth rate allows for higher plant throughput in a given production year, if this was combined with strains which produce higher titers of protein than those available in literature, it would allow E. coli to become competitive with CHO for the production of recombinant tPA. Experiments demonstrate that temperature control can be used to slow the metabolic rate of E. coli, allowing aerobic conditions to be maintained in the high density fermentations. Although temperature reduction has also been used to increase the yield of soluble protein, it is likely this occurs with reduced protein production. Temperature control was initiated using five minute moving averages to monitor overall oxygen and stirrer speed trends. Temperature was dropped 5 °C when averaged oxygen content fell below 18% and averaged stirrer speeds were greater than 1000 rpm. Temperature controlled runs for E. coli BL21DE3 producing eYFP appeared to allow the cultures to maintain better aerobic conditions. It is known that eYFP was produced since homogenized cell paste fluoresced yellow under UV light. However, protein analysis was hampered due to low protein production even after induction. Purifications involving large amounts of cell paste (50 g or more) were difficult to perform and all purificitiatons resulted in contamination by other proteins. Several recommendations can be made. The modeling would be greatly facilitated by additional information such as equipment specifications at large-scale production. The work with eYFP containing E. coli would be greatly enhanced by better strain selection. Choosing strains which over-express the protein of interest on the small scale would lead to better results in the fermentor. A densiometric analysis of the SDS PAGE gels run would allow a better understanding of general proteomic response to temperature control. When combined with mass spectrometry this may lead to different approaches in reducing temperature. Temperature control is often thought to increase soluble protein. From the densiometric SDS PAGE analysis of both the supernatant and pellet after homogenization it would be interesting to…

Subjects/Keywords: Modeling; Production; Recombinant Protein

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Yaeck, J. (2007). Production and Modeling of Recombinant Protein. (Thesis). University of Waterloo. Retrieved from http://hdl.handle.net/10012/2661

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Yaeck, Jason. “Production and Modeling of Recombinant Protein.” 2007. Thesis, University of Waterloo. Accessed January 24, 2021. http://hdl.handle.net/10012/2661.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Yaeck, Jason. “Production and Modeling of Recombinant Protein.” 2007. Web. 24 Jan 2021.

Vancouver:

Yaeck J. Production and Modeling of Recombinant Protein. [Internet] [Thesis]. University of Waterloo; 2007. [cited 2021 Jan 24]. Available from: http://hdl.handle.net/10012/2661.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Yaeck J. Production and Modeling of Recombinant Protein. [Thesis]. University of Waterloo; 2007. Available from: http://hdl.handle.net/10012/2661

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Boston University

22. Dehghani, Bijan. Development of an elastic sealant for surgical applications.

Degree: MS, Medical Sciences, 2015, Boston University

URL: http://hdl.handle.net/2144/16139

► The need to close wounds and prevent air/liquid leakage is commonly faced in surgical operations. It is a necessary step required for proper post-operative tissue… (more)

Subjects/Keywords: Biomedical engineering; Biomaterials; Elastin; Human recombinant protein; Sealant; Surgery; Tissue engineering

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Dehghani, B. (2015). Development of an elastic sealant for surgical applications. (Masters Thesis). Boston University. Retrieved from http://hdl.handle.net/2144/16139

Chicago Manual of Style (16^th Edition):

Dehghani, Bijan. “Development of an elastic sealant for surgical applications.” 2015. Masters Thesis, Boston University. Accessed January 24, 2021. http://hdl.handle.net/2144/16139.

MLA Handbook (7^th Edition):

Dehghani, Bijan. “Development of an elastic sealant for surgical applications.” 2015. Web. 24 Jan 2021.

Vancouver:

Dehghani B. Development of an elastic sealant for surgical applications. [Internet] [Masters thesis]. Boston University; 2015. [cited 2021 Jan 24]. Available from: http://hdl.handle.net/2144/16139.

Council of Science Editors:

Dehghani B. Development of an elastic sealant for surgical applications. [Masters Thesis]. Boston University; 2015. Available from: http://hdl.handle.net/2144/16139

University of Guelph

23. Hayward, Robin L. Production of Recombinant Human Butyrylcholinesterase in Nicotiana benthamiana.

Degree: MS, Department of Environmental Biology, 2012, University of Guelph

URL: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/4038

► Nerve agents (NAs) inhibit the essential enzyme acetylcholinesterase. Classified as chemical weapons, NAs are considered a threat to soldiers on the frontlines of warzones. Current… (more)

Subjects/Keywords: Butyrylcholinesterase; Nerve Agents; Organophosphates; Nicotiana benthamiana; Recombinant Protein

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Hayward, R. L. (2012). Production of Recombinant Human Butyrylcholinesterase in Nicotiana benthamiana. (Masters Thesis). University of Guelph. Retrieved from https://atrium.lib.uoguelph.ca/xmlui/handle/10214/4038

Chicago Manual of Style (16^th Edition):

Hayward, Robin L. “Production of Recombinant Human Butyrylcholinesterase in Nicotiana benthamiana.” 2012. Masters Thesis, University of Guelph. Accessed January 24, 2021. https://atrium.lib.uoguelph.ca/xmlui/handle/10214/4038.

MLA Handbook (7^th Edition):

Hayward, Robin L. “Production of Recombinant Human Butyrylcholinesterase in Nicotiana benthamiana.” 2012. Web. 24 Jan 2021.

Vancouver:

Hayward RL. Production of Recombinant Human Butyrylcholinesterase in Nicotiana benthamiana. [Internet] [Masters thesis]. University of Guelph; 2012. [cited 2021 Jan 24]. Available from: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/4038.

Council of Science Editors:

Hayward RL. Production of Recombinant Human Butyrylcholinesterase in Nicotiana benthamiana. [Masters Thesis]. University of Guelph; 2012. Available from: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/4038

24. Roux, Lauriane. Développement et validation d’un modèle cellulaire de kératopathie associée à l’aniridie : In vitro modeling of aniridia-related keratopathy and its validation.

Degree: Docteur es, Médecine. Biothérapies, 2018, Sorbonne Paris Cité

URL: http://www.theses.fr/2018USPCC276

►

L’aniridie est une pathologie rare principalement due à des mutations hétérozygotes sur PAX6, le gène contrôlant le développement oculaire et le maintien de l’homéostasie cornéenne.… (more)

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L’aniridie est une pathologie rare principalement due à des mutations hétérozygotes sur PAX6, le gène contrôlant le développement oculaire et le maintien de l’homéostasie cornéenne. Elle est caractérisée par une hypo/aplasie de l’iris, des atteintes de la rétine et du cristallin. La totalité des patients atteints développe aussi une kératopathie associée à l’aniridie (KAA), conduisant à une opacification progressive de leur cornée. La KAA a pour origine un déficit en cellules souches limbiques et diverses perturbations de l’épithélium et du stroma cornéens. Actuellement, il n’existe ni traitement pour soulager efficacement les patients, ni modèle cellulaire pour cette pathologie. Afin de pallier ces manques, le système CRISPR/Cas9 a été utilisé pour intégrer une mutation hétérozygote non-sens dans le gène PAX6 de cellules épithéliales limbiques immortalisées. Les cellules mutées présentent une expression réduite de PAX6 et une modulation de celle de ses gènes cibles, un ralentissement de la prolifération, de la clonogénicité et de la migration ainsi qu’une adhésion accrue. De plus, nous avons montré qu’un traitement de ces cellules avec une protéine recombinante PAX6, portant un peptide de pénétration cellulaire, permet une induction de l’expression endogène de PAX6 et également une restauration partielle du phénotype décrit précédemment. Cette réversion phénotypique valide le modèle d’haploinsuffisance développé et suggère l’implication de PAX6 dans les différentes fonctions restaurées. Les cellules mutées peuvent maintenant être utilisées pour le criblage d’outils thérapeutiques potentiels pour la KAA et pour d’autres défauts liés à une diminution du dosage de PAX6.

Aniridia is a rare panocular disease mainly due to PAX6 heterozygous mutations. PAX6 is the master gene of the eye development and it controls also the corneal homeostasis maintenance. Aniridia is characterized by an iris hypo/aplasia, retina and lens defects. All the aniridia patients will also develop an aniridia-related keratopathy (ARK) leading to a progressive corneal opacification. ARK is due to a limbal stem cell deficiency and alterations of corneal epithelium and stroma functions. Unfortunately, there is currently no efficient treatment to relief the patients and no cellular model for this pathology. To remedy these lacks, CRISPR/Cas9 system was used to insert a nonsense mutation into PAX6 gene of immortalized limbal epithelial cells. The mutated cells produce less PAX6 than the wild-type and PAX6 targets gene expression was modulated. They also display a marked slow-down proliferation, clonogenicity, migration and an enhanced adhesion. Moreover, we have shown that addition of recombinant PAX6 protein fused to a cell penetrating peptide to the culture medium was able to activate the endogenous PAX6 expression and to rescue some phenotypic defects of the mutated cells. Therefore, it validates the PAX6 haploinsufficiency model and suggests that PAX6 could be involved in all the rescued functions. The mutated cells can now be used to screen…

Advisors/Committee Members: Aberdam, Daniel (thesis director), Ferrigno, Olivier (thesis director).

Subjects/Keywords: Cellules épithéliales limbiques; Protéine recombinante; Limbal epithelial cells; Recombinant protein

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Roux, L. (2018). Développement et validation d’un modèle cellulaire de kératopathie associée à l’aniridie : In vitro modeling of aniridia-related keratopathy and its validation. (Doctoral Dissertation). Sorbonne Paris Cité. Retrieved from http://www.theses.fr/2018USPCC276

Chicago Manual of Style (16^th Edition):

Roux, Lauriane. “Développement et validation d’un modèle cellulaire de kératopathie associée à l’aniridie : In vitro modeling of aniridia-related keratopathy and its validation.” 2018. Doctoral Dissertation, Sorbonne Paris Cité. Accessed January 24, 2021. http://www.theses.fr/2018USPCC276.

MLA Handbook (7^th Edition):

Roux, Lauriane. “Développement et validation d’un modèle cellulaire de kératopathie associée à l’aniridie : In vitro modeling of aniridia-related keratopathy and its validation.” 2018. Web. 24 Jan 2021.

Vancouver:

Roux L. Développement et validation d’un modèle cellulaire de kératopathie associée à l’aniridie : In vitro modeling of aniridia-related keratopathy and its validation. [Internet] [Doctoral dissertation]. Sorbonne Paris Cité; 2018. [cited 2021 Jan 24]. Available from: http://www.theses.fr/2018USPCC276.

Council of Science Editors:

Roux L. Développement et validation d’un modèle cellulaire de kératopathie associée à l’aniridie : In vitro modeling of aniridia-related keratopathy and its validation. [Doctoral Dissertation]. Sorbonne Paris Cité; 2018. Available from: http://www.theses.fr/2018USPCC276

University of Waikato

25. Hardie, Sarah Elyse. Expression and Purification of Recombinant VMO1 Protein .

Degree: 2015, University of Waikato

URL: http://hdl.handle.net/10289/9609

► Hearing loss is a common sensory deficit that affects more than 275 million people worldwide. Identifying and understanding the genetic causes which underlie hearing loss… (more)

Subjects/Keywords: VMO1; Western blot; Antibodies; Recombinant protein; Hearing loss; pET system

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Hardie, S. E. (2015). Expression and Purification of Recombinant VMO1 Protein . (Masters Thesis). University of Waikato. Retrieved from http://hdl.handle.net/10289/9609

Chicago Manual of Style (16^th Edition):

Hardie, Sarah Elyse. “Expression and Purification of Recombinant VMO1 Protein .” 2015. Masters Thesis, University of Waikato. Accessed January 24, 2021. http://hdl.handle.net/10289/9609.

MLA Handbook (7^th Edition):

Hardie, Sarah Elyse. “Expression and Purification of Recombinant VMO1 Protein .” 2015. Web. 24 Jan 2021.

Vancouver:

Hardie SE. Expression and Purification of Recombinant VMO1 Protein . [Internet] [Masters thesis]. University of Waikato; 2015. [cited 2021 Jan 24]. Available from: http://hdl.handle.net/10289/9609.

Council of Science Editors:

Hardie SE. Expression and Purification of Recombinant VMO1 Protein . [Masters Thesis]. University of Waikato; 2015. Available from: http://hdl.handle.net/10289/9609

Universidade Nova

26. Silva, Maria Margarida dos Santos. Assessment of Notch1 ligands production - key protein targets in breast cancer.

Degree: 2014, Universidade Nova

URL: http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/13638

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Cell-to-cell communication is required for many biological processes in development and adult life. One of the most common systems utilized by a wide range of… (more)

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Cell-to-cell communication is required for many biological processes in development and adult life. One of the most common systems utilized by a wide range of eukaryotes is the Notch signalling pathway. Four Notch receptors and five ligands have been identified in mammals that interact via their extracellular domains leading to transcription activation. Studies have shown that the Notch ligands expression is undetectable in normal breast tissues, but moderate to high expression has been detected in breast cancer. Thus, any of the Notch1 ligands can be studied as possible therapeutic targets for breast cancer. To study Notch pathway proteins there is the need to obtain stable protein solutions. E. coli is the host of excellence for recombinant proteins for the ease of use, fast growth and high cell densities. However, the expression of mammalian proteins in such systems may overwhelm the bacterial cellular machinery, which does not possess the ability for post-translational modifications, or dedicated compartments for protein synthesis. Mammalian cells are therefore preferred, despite their technical and financial increased demands. We aim to determine the best expression and purification conditions for the different ligand protein constructs, to develop specific function-blocking antibodies using the Phage Display technology. Moreover, we propose to crystallize the Notch1 ligands alone and in complex with the phage display selected antibodies, unveiling molecular details. hJag2DE3 and hDll1DE6 proteins were purified from refolded inclusion bodies or mammalian cell culture supernatants, respectively, and purity was confirmed by SDS-PAGE (>95%). Protein produced in mammalian cells showed to be more stable, apparently with the physiological disulfide pattern, contrary to what was observed in the refolded protein. Several nano-scale crystallization experiments were set up in 96-well plates, but no positive result was obtained. We will continue to pursue for the best expression for the Notch ligand constructs in both expression systems.

Fundação para a Ciência e a Tecnologia - (PTDC/SAU-ONC/121670/2010), P-CUBE (Capacities Specific Programme Research Infrastructures, Project Number 227764) and NRS/LPCC “Terry Fox” 2013/2014 grants for funding, and iBET for hosting

Advisors/Committee Members: Bandeiras, Tiago, Barbas, Ana.

Subjects/Keywords: Notch signalling; Breast cancer; Recombinant protein expression; Macromolecular crystallisation

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Silva, M. M. d. S. (2014). Assessment of Notch1 ligands production - key protein targets in breast cancer. (Thesis). Universidade Nova. Retrieved from http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/13638

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Silva, Maria Margarida dos Santos. “Assessment of Notch1 ligands production - key protein targets in breast cancer.” 2014. Thesis, Universidade Nova. Accessed January 24, 2021. http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/13638.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Silva, Maria Margarida dos Santos. “Assessment of Notch1 ligands production - key protein targets in breast cancer.” 2014. Web. 24 Jan 2021.

Vancouver:

Silva MMdS. Assessment of Notch1 ligands production - key protein targets in breast cancer. [Internet] [Thesis]. Universidade Nova; 2014. [cited 2021 Jan 24]. Available from: http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/13638.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Silva MMdS. Assessment of Notch1 ligands production - key protein targets in breast cancer. [Thesis]. Universidade Nova; 2014. Available from: http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/13638

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Oxford

27. Rossi, Merja. Investigating cell type specific metabolism using GFP as a reporter protein.

Degree: PhD, 2015, University of Oxford

URL: https://ora.ox.ac.uk/objects/uuid:0c418362-63e7-496d-9ff6-584a0c54c127 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.711706

► Metabolic flux analysis (MFA) is a powerful technique for quantifying the intracellular fluxes in central carbon metabolism. It relies on detection of stable isotope labelling… (more)

▼ Metabolic flux analysis (MFA) is a powerful technique for quantifying the intracellular fluxes in central carbon metabolism. It relies on detection of stable isotope labelling from metabolites such as amino acids derived from protein. Current standard techniques are, however, unable to distinguish between different cell types in heterogeneous tissue. The aim of the thesis was to address this problem by developing and validating a strategy using green fluorescent protein (GFP) with cell type specific expression as a reporter protein for investigating the fluxes in specific cell types in the Arabidopsis thaliana root. The fundamental difficulty in applying a reporter protein strategy in a multicellular organism arises from the limited amount of recombinant protein expressed by the cells. The main novel contributions of the work in this thesis are threefold. First, a robust protocol for purification of GFP from the roots of Arabidopsis seedlings and for detection of reliable mass isotopomer distributions from the amino acids derived from GFP are described. Secondly, the reporter protein strategy is validated in this biological system with a focus on showing the data obtained by the use of the reporter protein is equal to that normally obtained from the total protein fraction. To expand on this, stable isotope labelling in isolated root hair cells is explored. These cells are easily isolated and show potential as a model system for cell type specific metabolism. Finally, the experimental data provide evidence for the feasibility of measuring data from specific cell types with appropriate mass spectrometric techniques. Analysis of cell type specific gene expression in this system suggests differences in the primary metabolism of different cell types cannot be ruled out without further investigation. Based on small scale in silico modelling described in this thesis, new solutions capable of providing data on sub-populations of cells are required, if central metabolism of the cell types differs significantly.

Subjects/Keywords: 572; Green fluorescent protein; Recombinant proteins; Arabidopsis thaliana

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APA (6^th Edition):

Rossi, M. (2015). Investigating cell type specific metabolism using GFP as a reporter protein. (Doctoral Dissertation). University of Oxford. Retrieved from https://ora.ox.ac.uk/objects/uuid:0c418362-63e7-496d-9ff6-584a0c54c127 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.711706

Chicago Manual of Style (16^th Edition):

Rossi, Merja. “Investigating cell type specific metabolism using GFP as a reporter protein.” 2015. Doctoral Dissertation, University of Oxford. Accessed January 24, 2021. https://ora.ox.ac.uk/objects/uuid:0c418362-63e7-496d-9ff6-584a0c54c127 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.711706.

MLA Handbook (7^th Edition):

Rossi, Merja. “Investigating cell type specific metabolism using GFP as a reporter protein.” 2015. Web. 24 Jan 2021.

Vancouver:

Rossi M. Investigating cell type specific metabolism using GFP as a reporter protein. [Internet] [Doctoral dissertation]. University of Oxford; 2015. [cited 2021 Jan 24]. Available from: https://ora.ox.ac.uk/objects/uuid:0c418362-63e7-496d-9ff6-584a0c54c127 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.711706.

Council of Science Editors:

Rossi M. Investigating cell type specific metabolism using GFP as a reporter protein. [Doctoral Dissertation]. University of Oxford; 2015. Available from: https://ora.ox.ac.uk/objects/uuid:0c418362-63e7-496d-9ff6-584a0c54c127 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.711706

University of Glasgow

28. Maloy, Kevin Joseph. The basis of the oral and parenteral adjuvant properties of immune stimulating complexes (ISCOMS).

Degree: PhD, 1996, University of Glasgow

URL: http://theses.gla.ac.uk/74948/ ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321045

► There is currently a great deal of interest in the development of vaccines using purified recombinant protein antigens. For practical and scientific reasons, it would… (more)

▼ There is currently a great deal of interest in the development of vaccines using purified recombinant protein antigens. For practical and scientific reasons, it would be advantageous if these vaccines could be administered orally. This is esssential for stimulating widespread immunity at the mucosal surfaces where most pathogens are encountered and, as the success of the oral polio vaccine shows, this can also protect against systemic infection. Furthermore, oral immunisation is one of the few effective means allowing passive transfer of immunity from mother to infant via milk. The poor immunogenicity of purified soluble proteins, their inability to induce class I MHC-restricted CTL responses and the systemic tolerance induced by their oral administration are major obstacles to the development of synthetic oral vaccines. Thus, there is a need for well characterised vaccine vectors which will circumvent these difficulties and allow soluble protein antigens to be immunogenic when administered orally. Immune stimulating complexes (ISCOMS) are lipophilic, cage-like particles composed of cholesterol, phospholipid and the saponin adjuvant Quil A. Parenteral immunisation with ISCOMS is known to induce humoral and cell-mediated immune responses as well as protective immunity against a number of infections. However, their efficacy by mucosal routes and their use with non-hydrophobic proteins had received little attention. My studies exploited a recently established method for incorporating purified OVA into ISCOMS and aimed to extend preliminary work in the laboratory showing that ISCOMS-OVA were immunogenic by oral and parenteral routes. I therefore explored the full range of responses induced and investigated the immunological basis of the adjuvant properties of ISCOMS when used by the oral and parenteral routes. A single parenteral immunisation with as little as 5-10mug ISCOMS-OVA primed potent OVA-specific systemic DTH and IgG responses and high levels of class I MHC-restricted OVA-specific CTL activity were detected in the spleen. These findings indicate that ISCOMS allow presentation of incorporated protein to the class II MHC-restricted T cells required for antibody and DTH responses, as well as permitting efficient entry of the protein into the endogenous antigen processing pathway required for stimulation of class 1 MHC-restricted CTL responses. A multiple dose oral immunisation schedule induced the same range of systemic immune responses as parenteral immunisation and also stimulated the production of secretory IgA antibodies in the intestine and local CTL precursors in MLN. These findings indicate that ISCOMS may allow protein antigen to subvert the regulatory mechanisms which normally cause protein antigens to induce tolerance at mucosal surfaces. One of the most novel properties of ISCOMS was their ability to stimulate potent class I MHC-restricted CTL responses to exogenous protein antigens in vivo. As this would be important for a vaccine capable of stimulating protection against many viral infections, I…

Subjects/Keywords: 615.1; Vaccines: Recombinant protein antigens

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Maloy, K. J. (1996). The basis of the oral and parenteral adjuvant properties of immune stimulating complexes (ISCOMS). (Doctoral Dissertation). University of Glasgow. Retrieved from http://theses.gla.ac.uk/74948/ ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321045

Chicago Manual of Style (16^th Edition):

Maloy, Kevin Joseph. “The basis of the oral and parenteral adjuvant properties of immune stimulating complexes (ISCOMS).” 1996. Doctoral Dissertation, University of Glasgow. Accessed January 24, 2021. http://theses.gla.ac.uk/74948/ ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321045.

MLA Handbook (7^th Edition):

Maloy, Kevin Joseph. “The basis of the oral and parenteral adjuvant properties of immune stimulating complexes (ISCOMS).” 1996. Web. 24 Jan 2021.

Vancouver:

Maloy KJ. The basis of the oral and parenteral adjuvant properties of immune stimulating complexes (ISCOMS). [Internet] [Doctoral dissertation]. University of Glasgow; 1996. [cited 2021 Jan 24]. Available from: http://theses.gla.ac.uk/74948/ ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321045.

Council of Science Editors:

Maloy KJ. The basis of the oral and parenteral adjuvant properties of immune stimulating complexes (ISCOMS). [Doctoral Dissertation]. University of Glasgow; 1996. Available from: http://theses.gla.ac.uk/74948/ ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321045

University of Manchester

29. Idicula Babu, Benoy. Epigallocatechin-3-gallate and recombinant human activated protein C and the modulation of acute pancreatitis.

Degree: Thesis (M.D.), 2012, University of Manchester

URL: https://www.research.manchester.ac.uk/portal/en/theses/epigallocatechin3gallate-and-recombinant-human-activated-protein-c-and-the-modulation-of-acute-pancreatitis(5fc797e9-cf5b-4aab-841d-62c3a8cd82c6).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.566553

► Effective management of acute pancreatitis has for centuries eluded mankind. The disease has a wide spectrum of presentation; the milder form is usually a self… (more)

▼ Effective management of acute pancreatitis has for centuries eluded mankind. The disease has a wide spectrum of presentation; the milder form is usually a self limiting condition, whereas the severe form presents as a highly morbid and frequently lethal attack. The ability to predict disease progression on admission would aid in the comprehensive and multidisciplinary management of patients. The perfect predictor of disease progression has been an elusive factor hindering the management of the disease. On systematically reviewing literature and identifying appropriate biochemical markers in predicting progression of acute pancreatitis, the ideal predictor would be a combination of biochemical, clinical and contemporary organ dysfunction scoring systems. Early prediction of disease progression however, is important in the better management of the disease. The pathophysiological changes of acinar cell injury and death are the earliest events that occur in acute pancreatitis. Identification of potential pharmacological interventions offered through valuable insight in to experimental and clinical acute pancreatitis may lead on to the development of various natural and synthetic potential disease modifiers. Green Tea Extracts (GTE) consumed in many parts of the world has been examined as a potential therapeutic medication. Experimental results have demonstrated the effect of GTE on the oxidative pathway significantly ameliorating the effects of pancreatic injury. The various green tea catechins especially Epigallocatechin-3- gallate (EGCG) can perhaps be useful lead compounds for new drug discovery. With no specific targeted therapy for severe acute pancreatitis at present, various medications have been tested. The possibility of targeting initial acinar cell injury may not be a feasible option as patient presentation and management would usually be after this phase. As the disease progresses, severe acute pancreatitis is characterised by inflammation and necrosis. The hypothesis of preserving pancreatic parenchymal microvascular patency and thus ameliorating pancreatic injury through the early administration of recombinant human Activated Protein C (rhAPC) has identified a potential treatment for acute pancreatitis. rhAPC converted from its inactive precursor, protein C, by thrombin acts through fibrinolysis and inhibition of thrombosis. Studies on rhAPC in experimental acute pancreatitis examined the modulation of rhAPC on inflammatory markers, morphology, microvascular thrombosis and apoptosis. The encouraging results from initial experimental work helped set up the Phase 2 clinical trial of administering rhAPC early on in severe acute pancreatitis. Prior to taking this significant step from bench to bed side, the variation in functional protein C levels with the severity of the disease was examined as a precursor to the Phase 2 trial.

Subjects/Keywords: 616.3; Green tea extracts; Recombinant human activated protein C; Acute pancreatitis

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Idicula Babu, B. (2012). Epigallocatechin-3-gallate and recombinant human activated protein C and the modulation of acute pancreatitis. (Doctoral Dissertation). University of Manchester. Retrieved from https://www.research.manchester.ac.uk/portal/en/theses/epigallocatechin3gallate-and-recombinant-human-activated-protein-c-and-the-modulation-of-acute-pancreatitis(5fc797e9-cf5b-4aab-841d-62c3a8cd82c6).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.566553

Chicago Manual of Style (16^th Edition):

Idicula Babu, Benoy. “Epigallocatechin-3-gallate and recombinant human activated protein C and the modulation of acute pancreatitis.” 2012. Doctoral Dissertation, University of Manchester. Accessed January 24, 2021. https://www.research.manchester.ac.uk/portal/en/theses/epigallocatechin3gallate-and-recombinant-human-activated-protein-c-and-the-modulation-of-acute-pancreatitis(5fc797e9-cf5b-4aab-841d-62c3a8cd82c6).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.566553.

MLA Handbook (7^th Edition):

Idicula Babu, Benoy. “Epigallocatechin-3-gallate and recombinant human activated protein C and the modulation of acute pancreatitis.” 2012. Web. 24 Jan 2021.

Vancouver:

Idicula Babu B. Epigallocatechin-3-gallate and recombinant human activated protein C and the modulation of acute pancreatitis. [Internet] [Doctoral dissertation]. University of Manchester; 2012. [cited 2021 Jan 24]. Available from: https://www.research.manchester.ac.uk/portal/en/theses/epigallocatechin3gallate-and-recombinant-human-activated-protein-c-and-the-modulation-of-acute-pancreatitis(5fc797e9-cf5b-4aab-841d-62c3a8cd82c6).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.566553.

Council of Science Editors:

Idicula Babu B. Epigallocatechin-3-gallate and recombinant human activated protein C and the modulation of acute pancreatitis. [Doctoral Dissertation]. University of Manchester; 2012. Available from: https://www.research.manchester.ac.uk/portal/en/theses/epigallocatechin3gallate-and-recombinant-human-activated-protein-c-and-the-modulation-of-acute-pancreatitis(5fc797e9-cf5b-4aab-841d-62c3a8cd82c6).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.566553

Univerzitet u Beogradu

30. Bašica, Branka, 1985-, 59524617. Филогенетска анализа гена, развојна, ткивна и полна дистрибуција глутатион-ѕ-трансфераза рибе зебрице - функционална карактеризација одабраних рекомбинантних протеина.

Degree: Biološki fakultet, 2020, Univerzitet u Beogradu

URL: https://fedorabg.bg.ac.rs/fedora/get/o:22978/bdef:Content/get

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Биологија - Молекуларна биологија, Физиологија / Biology - Molecular biology, Physiology

Глутатион-S-трансферазе (GST) припадају вишефункционалној суперпородици ензима и имају разноврсне каталитичке улоге у организму, са… (more)

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Биологија - Молекуларна биологија, Физиологија / Biology - Molecular biology, Physiology

Глутатион-S-трансферазе (GST) припадају вишефункционалној суперпородици ензима и имају разноврсне каталитичке улоге у организму, са основном функцијом конјугације редукованог глутатиона са разноврсним ксенобиотицима у циљу смањења њихове токсичности. Улога Gst у процесу детоксификације код риба је препозната током 1970-их година, али према доступној литератури, само је неколико рибљих ензима Gst функционално окарактерисано. У овој дисертацији урађена је свеобухватна карактеризација ензимске суперпородице Gst зебрица (лат. Danio rerio), важном кичмењачком модел организму, са циљем бољег разумевања диверзификације и еволуције, физиолошког значаја и улоге ензима Gst као биомаркера изложености одређеним хемикалијама. Комбинацијом биоинформатичких метода, анализом експримирања гена, експримирањем и пречишћавањем рекомбинтантних протеина и применом ензимских тестова и тестирањем инхибиторног потенцијала различитих супстанци урађена је филогенетска анализа, развојна, ткивна и полна дистрибуција иРНК гена gst зебрице и функционална карактеризација одабраних рекомбинантних протеина (Gstr1, Gstt2 и Gstm3). Филогенетска анализа је открила 27 чланова суперпородице gst у геному зебрице, подељених у 9 класа. Развојнa, ткивнa и полнa дистрибуција иРНК, као и функционална сличност са хуманим ортолозима, указујe да су представници класа Pi, Theta, Zeta и Rho значајни у процесима биотрансформације ксенобиотика, и да потенцијално могу имати кључну улогу у одређеним физиолошким процесима. Резултати указују на могућу улогу Gstr1 у стероидогенези, метаболизму и/или физиолошкој активности андрогена код риба, насупрот изостанку овакве функције према естрогенима, као и заштити риба од штетних загађивача из спољашње средине, као што су органофосфорни инсектициди и лекови.

Advisors/Committee Members: Kovačević, Radmila, 1948-, 13149799.

Subjects/Keywords: enzymatic activity; functional characterization; Gst; mRNA distribution; phylogeny; recombinant protein; zebrafish

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APA (6^th Edition):

Bašica, Branka, 1985-, 5. (2020). Филогенетска анализа гена, развојна, ткивна и полна дистрибуција глутатион-ѕ-трансфераза рибе зебрице - функционална карактеризација одабраних рекомбинантних протеина. (Thesis). Univerzitet u Beogradu. Retrieved from https://fedorabg.bg.ac.rs/fedora/get/o:22978/bdef:Content/get

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Bašica, Branka, 1985-, 59524617. “Филогенетска анализа гена, развојна, ткивна и полна дистрибуција глутатион-ѕ-трансфераза рибе зебрице - функционална карактеризација одабраних рекомбинантних протеина.” 2020. Thesis, Univerzitet u Beogradu. Accessed January 24, 2021. https://fedorabg.bg.ac.rs/fedora/get/o:22978/bdef:Content/get.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Bašica, Branka, 1985-, 59524617. “Филогенетска анализа гена, развојна, ткивна и полна дистрибуција глутатион-ѕ-трансфераза рибе зебрице - функционална карактеризација одабраних рекомбинантних протеина.” 2020. Web. 24 Jan 2021.

Vancouver:

Bašica, Branka, 1985- 5. Филогенетска анализа гена, развојна, ткивна и полна дистрибуција глутатион-ѕ-трансфераза рибе зебрице - функционална карактеризација одабраних рекомбинантних протеина. [Internet] [Thesis]. Univerzitet u Beogradu; 2020. [cited 2021 Jan 24]. Available from: https://fedorabg.bg.ac.rs/fedora/get/o:22978/bdef:Content/get.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Bašica, Branka, 1985- 5. Филогенетска анализа гена, развојна, ткивна и полна дистрибуција глутатион-ѕ-трансфераза рибе зебрице - функционална карактеризација одабраних рекомбинантних протеина. [Thesis]. Univerzitet u Beogradu; 2020. Available from: https://fedorabg.bg.ac.rs/fedora/get/o:22978/bdef:Content/get

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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