subject:(Recombinant human insulin)

University of Vermont

1. Qian, Xi. Regulation of β-Casein Gene Expression by Octamer Transcription Factors and Utilization of β-Casein Gene Promoter to Produce Recombinant Human Proinsulin in the Transgenic Milk.

Degree: PhD, Animal Science, 2014, University of Vermont

URL: https://scholarworks.uvm.edu/graddis/316

► β-Casein is a major milk protein, which is synthesized in mammary alveolar secretory epithelial cells (MECs) upon the stimulation of lactogenic hormones, mainly prolactin… (more)

▼ β-Casein is a major milk protein, which is synthesized in mammary alveolar secretory epithelial cells (MECs) upon the stimulation of lactogenic hormones, mainly prolactin and glucocorticoids (HP). Previous studies revealed that the proximal promoter (-258 bp to +7 bp) of the β-casein gene is sufficient for induction of the promoter activity by HP. This proximal region contains the binding sites for the signal transducer and activator of transcription 5 (STAT5), glucocorticoid receptor (GR), and octamer transcription factors (Oct). STAT5 and GR are essential downstream mediators of prolactin and glucocorticoid signaling, respectively. This study investigated the functions of Oct-1 and Oct-2 in HP induction of β-casein gene expression. By transiently transfection experiment, we showed that individual overexpression of Oct-1 and Oct-2 further enhanced HP-induced β-casein promoter activity, respectively, while Oct-1 and Oct-2 knockdown significantly inhibited the HP-induced β-casein promoter activity, respectively. HP rapidly induced the binding of both Oct-1 and Oct-2 to the β-casein promoter, and this induction was not mediated by either increasing their expression or inducing their translocation to the nucleus. In MECs, Oct-2 was found to physically interact with Oct-1 regardless of HP treatment. However, HP induced physical interactions of Oct-1 or Oct-2 with both STAT5 and GR. Although the interaction between Oct-1 and Oct-2 did not synergistically stimulate HP-induced β-casein gene promoter activity, the synergistic effect was observed for the interactions of Oct-1 or Oct-2 with STAT5 and GR. The interactions of Oct-1 with STAT5 and GR enhanced or stabilized the binding of STAT5 and GR to the promoter. Abolishing the interaction between Oct-1 and STAT5 significantly reduced the hormonal induction of β-casein gene transcription. Thus, our study indicates that HP activate β-casein gene expression by inducing the physical interactions of Oct-1 and Oct-2 with STAT5 and GR in mouse MECs. There is a high and increasing demand for insulin because of the rapid increase in diabetes incidence worldwide. However, the current manufacturing capacities can barely meet the increasing global demand for insulin, and the cost of insulin production keeps rising. The mammary glands of dairy animals have been regarded as ideal bioreactors for mass production of therapeutically important human proteins. We tested the feasibility of producing human proinsulin in the milk of transgenic mice. In this study, four lines of transgenic mice were generated to harbor the human insulin gene driven by the goat β-casein gene promoter. The recombinant human proinsulin was detected in the milk by Western blotting and enzyme-linked immunosorbent assay. The highest expression level of human proinsulin was as high as 8.1 μg/µl in milk of transgenic mice at mid-lactation. The expression of the transgene was only detected in the mammary gland during lactation. The transgene expression profile throughout lactation resembled the milk yield curve,… Advisors/Committee Members: Feng-Qi Zhao.

Subjects/Keywords: Bioreactor; Hormonal regulation; Milk protein; Octamer binding transcription factor; Recombinant human insulin; Transcriptional regulation; Biochemistry; Biomedical; Molecular Biology

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APA (6^th Edition):

Qian, X. (2014). Regulation of β-Casein Gene Expression by Octamer Transcription Factors and Utilization of β-Casein Gene Promoter to Produce Recombinant Human Proinsulin in the Transgenic Milk. (Doctoral Dissertation). University of Vermont. Retrieved from https://scholarworks.uvm.edu/graddis/316

Chicago Manual of Style (16^th Edition):

Qian, Xi. “Regulation of β-Casein Gene Expression by Octamer Transcription Factors and Utilization of β-Casein Gene Promoter to Produce Recombinant Human Proinsulin in the Transgenic Milk.” 2014. Doctoral Dissertation, University of Vermont. Accessed January 27, 2021. https://scholarworks.uvm.edu/graddis/316.

MLA Handbook (7^th Edition):

Qian, Xi. “Regulation of β-Casein Gene Expression by Octamer Transcription Factors and Utilization of β-Casein Gene Promoter to Produce Recombinant Human Proinsulin in the Transgenic Milk.” 2014. Web. 27 Jan 2021.

Vancouver:

Qian X. Regulation of β-Casein Gene Expression by Octamer Transcription Factors and Utilization of β-Casein Gene Promoter to Produce Recombinant Human Proinsulin in the Transgenic Milk. [Internet] [Doctoral dissertation]. University of Vermont; 2014. [cited 2021 Jan 27]. Available from: https://scholarworks.uvm.edu/graddis/316.

Council of Science Editors:

Qian X. Regulation of β-Casein Gene Expression by Octamer Transcription Factors and Utilization of β-Casein Gene Promoter to Produce Recombinant Human Proinsulin in the Transgenic Milk. [Doctoral Dissertation]. University of Vermont; 2014. Available from: https://scholarworks.uvm.edu/graddis/316

Georgia Tech

2. Tang, Shiue-Cheng. Genetic engineering of non-beta-cells for regulated insulin secretion.

Degree: PhD, Chemical Engineering, 2003, Georgia Tech

URL: http://hdl.handle.net/1853/5375

Subjects/Keywords: Recombinant human insulin; Pancreatic beta cells; Diabetes; Recombinant human insulin; Pancreatic beta cells; Diabetes

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

APA (6^th Edition):

Tang, S. (2003). Genetic engineering of non-beta-cells for regulated insulin secretion. (Doctoral Dissertation). Georgia Tech. Retrieved from http://hdl.handle.net/1853/5375

Chicago Manual of Style (16^th Edition):

Tang, Shiue-Cheng. “Genetic engineering of non-beta-cells for regulated insulin secretion.” 2003. Doctoral Dissertation, Georgia Tech. Accessed January 27, 2021. http://hdl.handle.net/1853/5375.

MLA Handbook (7^th Edition):

Tang, Shiue-Cheng. “Genetic engineering of non-beta-cells for regulated insulin secretion.” 2003. Web. 27 Jan 2021.

Vancouver:

Tang S. Genetic engineering of non-beta-cells for regulated insulin secretion. [Internet] [Doctoral dissertation]. Georgia Tech; 2003. [cited 2021 Jan 27]. Available from: http://hdl.handle.net/1853/5375.

Council of Science Editors:

Tang S. Genetic engineering of non-beta-cells for regulated insulin secretion. [Doctoral Dissertation]. Georgia Tech; 2003. Available from: http://hdl.handle.net/1853/5375

3. Nicolau Brito da Cunha. Expressão de cinco diferentes proteínas de interesse farmacológico em sementes transgênicas de soja [Glycine Max L. (Merril)].

Degree: 2008, Universidade Católica de Brasilia

URL: http://www.bdtd.ucb.br/tede/tde_busca/arquivo.php?codArquivo=708

►

As plantas superiores representam um sistema conveniente e versátil para a produção em larga escala de proteínas heterólogas. A confirmação desse potencial tem sido demonstrada… (more)

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As plantas superiores representam um sistema conveniente e versátil para a produção em larga escala de proteínas heterólogas. A confirmação desse potencial tem sido demonstrada pelo sucesso comercial de sistemas de produção desses polipeptídeos, baseados na transformação genética das mais variadas espécies vegetais, resultando atualmente na produção eficiente de mais de 100 diferentes proteínas heterólogas. Uma porcentagem significativa desse conjunto de polipeptídeos recombinantes engloba aquelas cujo interesse farmacológico e terapêutico se mostra evidenciado em função das propriedades bioquímicas apresentadas por essas moléculas, tais como hormônios, fatores de crescimento, anticorpos e fatores de coagulação sangüinea. Biorreatores vegetais apresentam inúmeras vantagens em relação a outros sistemas de produção de biofármacos recombinantes, principalmente no que diz respeito à praticidade, economia e segurança, apesar de algumas limitações envolvendo homogeneidade do produto, baixa estabilidade protéica e baixos níveis de expressão ainda se mostrarem recorrentes. Dessa maneira a escolha de plantas de soja [Glycine max L.(Merril)] como veículos de expressão de biofármacos de interesse comercial visa não apenas contornar algumas dessas dificuldades como também aproveitar características extremamente favoráveis para sua utilização como biorreatores de fármacos protéicos, tais como sua baixa freqüência de alogamia, ausência de parentes silvestres no Brasil e seu potencial natural de acúmulo protéico nas sementes, através de uma estratégia de expressão tecido-específico e de endereçamento molecular desses polipeptídeos para prolongamentos dos retículos endoplasmáticos de células cotiledonares das sementes, denominados corpos protéicos. Para tanto, foram construídos diferentes cassetes de expressão em nosso laboratório contendo cinco seqüências codificadoras de biofármacos com atividade efetora em humanos e de considerável impacto social: a pró-insulina, o hormônio de crescimento e o fator IX de coagulação sangüínea humanos; e fragmentos de anticorpos anti-câncer de mama e anti-reperfusão pósisquêmica, sob controle do promotor tecido-específico de sementes da β-faseolina do feijoeiro-comum (Phaseolus vulgaris L.) ― a mais abundante nas sementes de feijão; além do peptídeo-sinal da α-coixina de Coix (Coix lacrima-jobi L.), responsável pelo endereçamento dos polipeptídeos para os corpos protéicos cotiledonares. Após a clonagem dos cassetes de expressão foram obtidas diferentes linhagens transgênicas de soja através do processo de biobalística e análises moleculares e bioquímicas foram realizadas para avaliar o número de cópias dos diferentes transgenes integradas ao genoma de cada linhagem, a segregação dos transgenes em sucessivas gerações de multiplicação de sementes em casa de vegetação, os níveis de expressão dos transgenes, a cinética química de expressão ao longo do ciclo de produção de sementes e a localização subcelular do acúmulo de proteínas heterólogas nas sementes.

Superior plants represent a…

Advisors/Committee Members: Natália Cristina Verza Ferreira, Octavio Luiz Franco, Elibio Leopoldo Rech Filho.

Subjects/Keywords: anticorpos recombinantes; fator de coagulação sangüínea; hormônio de crescimento humano; insulina humana; biorreatores vegetais; proteínas heterólogas; soja transgênica; soja, soja melhoramento genético; insulina; hormônios; Ciências Biológicas; plant-based bioreactors; heterologous proteins; transgenic soybean; human insulin; human growth hormone; coagulation factor; recombinant antibodies; Ciências Biológicas

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

APA (6^th Edition):

Cunha, N. B. d. (2008). Expressão de cinco diferentes proteínas de interesse farmacológico em sementes transgênicas de soja [Glycine Max L. (Merril)]. (Masters Thesis). Universidade Católica de Brasilia. Retrieved from http://www.bdtd.ucb.br/tede/tde_busca/arquivo.php?codArquivo=708

Chicago Manual of Style (16^th Edition):

Cunha, Nicolau Brito da. “Expressão de cinco diferentes proteínas de interesse farmacológico em sementes transgênicas de soja [Glycine Max L. (Merril)].” 2008. Masters Thesis, Universidade Católica de Brasilia. Accessed January 27, 2021. http://www.bdtd.ucb.br/tede/tde_busca/arquivo.php?codArquivo=708.

MLA Handbook (7^th Edition):

Cunha, Nicolau Brito da. “Expressão de cinco diferentes proteínas de interesse farmacológico em sementes transgênicas de soja [Glycine Max L. (Merril)].” 2008. Web. 27 Jan 2021.

Vancouver:

Cunha NBd. Expressão de cinco diferentes proteínas de interesse farmacológico em sementes transgênicas de soja [Glycine Max L. (Merril)]. [Internet] [Masters thesis]. Universidade Católica de Brasilia; 2008. [cited 2021 Jan 27]. Available from: http://www.bdtd.ucb.br/tede/tde_busca/arquivo.php?codArquivo=708.

Council of Science Editors:

Cunha NBd. Expressão de cinco diferentes proteínas de interesse farmacológico em sementes transgênicas de soja [Glycine Max L. (Merril)]. [Masters Thesis]. Universidade Católica de Brasilia; 2008. Available from: http://www.bdtd.ucb.br/tede/tde_busca/arquivo.php?codArquivo=708

4. Gulino, Angela Marie. Insulin secretion dynamics of recombinant hepatic and intestinal cells.

Degree: MS, Biomedical Engineering, 2008, Georgia Tech

URL: http://hdl.handle.net/1853/28220

► Hepatic and intestinal endocrine cells are potentially helpful targets for recombinant insulin expression. As the two cell types exhibit different secretion kinetics,it has been hypothesized… (more)

▼ Hepatic and intestinal endocrine cells are potentially helpful targets for recombinant insulin expression. As the two cell types exhibit different secretion kinetics,it has been hypothesized that a combination of the two would better approximate insulin secretion kinetics from normal, functioning beta-cells than either cell type alone. This hypothesis was tested using two hepatic cell lines transiently transduced with one of three adenoviruses for insulin expression along with a stably transfected recombinant intestinal L cell line. The insulin secretion kinetics were analyzed for both the hepatic and intestinal cells to determine the potential of combining them to reproduce the insulin secretion kinetics of a normal, functioning beta-cell. It was observed that the two recombinant hepatic cell lines secreted insulin in a more sustained manner exhibiting slower release kinetics. They also exhibited an increase in insulin secretion when stimulated by the cocktail of nutrient secretagogues (glucose and meat hydrolysate) versus stimulating with only glucose. The cells transduced with the adenovirus containing an additional cytomegalovirus (CMV) promoter and green fluorescent protein (GFP) exhibited the highest insulin secretion after stimulation, whereas the cells transduced with an adenovirus encoding for destabilized preproinsulin mRNA exhibited the lowest secretion rates. The recombinant intestinal cell line (GLUTag-INS) secreted insulin with rapid kinetics upon stimulation, apparently due to the presence of secretory granules containing pre-synthesized insulin. The experiments demonstrated that the cells stimulated with medium containing only meat hydrolysate exhibited a significantly higher insulin secretion relative to secretagogue-free controls. The insulin secretion was not further enhanced when meat hydrolysate was combined with glucose. Advisors/Committee Members: Dr. Athanassios Sambanis (Committee Chair), Dr. Barbara Boyan (Committee Member), Dr. Peter Thule (Committee Member).

Subjects/Keywords: Genetic engineering; Diabetes; Liver; Insulin; Kinetics; Intestinal; Liver cells; Intestines; Endocrinology; Recombinant human insulin; Pancreatic beta cells

…type 2 hB10 human B10 modified IDD insulin-dependent diabetes Kb kilobases MH meat… …potentially helpful targets for recombinant insulin expression. As the two cell types exhibit… …of three adenoviruses for insulin expression along with a stably transfected recombinant… …objective is to analyze insulin secretion kinetics of recombinant hepatic and intestinal cell… …the insulin. Furin modified human insulin cDNA has been shown to lower blood glucose levels…

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

APA (6^th Edition):

Gulino, A. M. (2008). Insulin secretion dynamics of recombinant hepatic and intestinal cells. (Masters Thesis). Georgia Tech. Retrieved from http://hdl.handle.net/1853/28220

Chicago Manual of Style (16^th Edition):

Gulino, Angela Marie. “Insulin secretion dynamics of recombinant hepatic and intestinal cells.” 2008. Masters Thesis, Georgia Tech. Accessed January 27, 2021. http://hdl.handle.net/1853/28220.

MLA Handbook (7^th Edition):

Gulino, Angela Marie. “Insulin secretion dynamics of recombinant hepatic and intestinal cells.” 2008. Web. 27 Jan 2021.

Vancouver:

Gulino AM. Insulin secretion dynamics of recombinant hepatic and intestinal cells. [Internet] [Masters thesis]. Georgia Tech; 2008. [cited 2021 Jan 27]. Available from: http://hdl.handle.net/1853/28220.

Council of Science Editors:

Gulino AM. Insulin secretion dynamics of recombinant hepatic and intestinal cells. [Masters Thesis]. Georgia Tech; 2008. Available from: http://hdl.handle.net/1853/28220

University of Adelaide

5. van Renen, Margaret Jean. Effects of GH on the IGFs and IGFBPs in children with chronic renal failure and transplantation.

Degree: 1996, University of Adelaide

URL: http://hdl.handle.net/2440/38447

► This thesis involves the retrospective investigation of the insulin-like growth factors and their binding proteins in the serum of children with chronic renal failure (CRF)… (more)

Subjects/Keywords: Chronic renal failure in children; Recombinant human somatotropin; Somatomedin; Insulin-like growth factor-binding proteins; Physiology, Pathological; .

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

APA (6^th Edition):

van Renen, M. J. (1996). Effects of GH on the IGFs and IGFBPs in children with chronic renal failure and transplantation. (Thesis). University of Adelaide. Retrieved from http://hdl.handle.net/2440/38447

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):

van Renen, Margaret Jean. “Effects of GH on the IGFs and IGFBPs in children with chronic renal failure and transplantation.” 1996. Thesis, University of Adelaide. Accessed January 27, 2021. http://hdl.handle.net/2440/38447.

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

MLA Handbook (7^th Edition):

van Renen, Margaret Jean. “Effects of GH on the IGFs and IGFBPs in children with chronic renal failure and transplantation.” 1996. Web. 27 Jan 2021.

Vancouver:

van Renen MJ. Effects of GH on the IGFs and IGFBPs in children with chronic renal failure and transplantation. [Internet] [Thesis]. University of Adelaide; 1996. [cited 2021 Jan 27]. Available from: http://hdl.handle.net/2440/38447.

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

Council of Science Editors:

van Renen MJ. Effects of GH on the IGFs and IGFBPs in children with chronic renal failure and transplantation. [Thesis]. University of Adelaide; 1996. Available from: http://hdl.handle.net/2440/38447

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

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