subject:(fluorescent protein)

University of Alberta

1. Alford, Spencer Caleb. Development of fluorogenic fluorescent protein heterodimers.

Degree: PhD, Department of Chemistry, 2012, University of Alberta

URL: https://era.library.ualberta.ca/files/hh63sw19n

► Fluorescent proteins (FPs) are indispensible biochemical tools. The concerted efforts of protein engineers have produced FPs spanning the visible colour spectrum. This wide variety of… (more)

▼ Fluorescent proteins (FPs) are indispensible biochemical tools. The concerted efforts of protein engineers have produced FPs spanning the visible colour spectrum. This wide variety of FPs has greatly facilitated the development of FP-based biosensors. However, researchers rely on relatively few fundamental biosensor design templates. Förster resonance energy transfer and bimolecular complementation are the principal FP-based technologies for live cell imaging of physiological events, such as changes in small molecule concentration, enzymatic activities, and protein-protein interactions. Although widely used, these techniques are often restrictive due to poor signal-to-noise ratios and irreversible sensing, respectively. Furthermore, examples of these biosensor strategies incorporating red FPs are limited. In this thesis we describe our efforts to address this shortcoming in the area of FP-based biosensors. We developed a dimerization-dependent red FP (ddRFP) that serves as an alternative template for biosensor construction. The prototype ddRFP was engineered from a homodimeric variant of a Discosoma red FP. Through extensive directed evolution the homodimer was converted into a fluorogenic obligate heterodimer. The reversible changes in fluorescence intensity that result from association of the ddRFP monomeric constituents, or the irreversible decrease that accompanies dissociation of covalently linked partners following linker cleavage, provides a useful spectroscopic signal for biosensing applications. Specifically, we demonstrated that ddRFP is useful for detecting in vitro protein-protein interactions, as well as imaging changes in calcium ion concentration and activation of caspase-3 in live cells. We also report the expansion of the ddFP colour palette through the development of green (ddGFP) and yellow (ddYFP) ddFP variants. These variants have several improvements relative to the ddRFP prototype including increased in vitro contrast and brightness for ddGFP, and a reduced pKa for ddYFP. While their utility for some live cell imaging applications is restricted due to low dissociation constants, ddGFP proved to be a useful fluorescent label of intermembrane contact sites between the endoplasmic reticulum and the mitochondrial network.

Subjects/Keywords: protein engineering; fluorescent protein; biosensor

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Alford, S. C. (2012). Development of fluorogenic fluorescent protein heterodimers. (Doctoral Dissertation). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/hh63sw19n

Chicago Manual of Style (16^th Edition):

Alford, Spencer Caleb. “Development of fluorogenic fluorescent protein heterodimers.” 2012. Doctoral Dissertation, University of Alberta. Accessed April 11, 2021. https://era.library.ualberta.ca/files/hh63sw19n.

MLA Handbook (7^th Edition):

Alford, Spencer Caleb. “Development of fluorogenic fluorescent protein heterodimers.” 2012. Web. 11 Apr 2021.

Vancouver:

Alford SC. Development of fluorogenic fluorescent protein heterodimers. [Internet] [Doctoral dissertation]. University of Alberta; 2012. [cited 2021 Apr 11]. Available from: https://era.library.ualberta.ca/files/hh63sw19n.

Council of Science Editors:

Alford SC. Development of fluorogenic fluorescent protein heterodimers. [Doctoral Dissertation]. University of Alberta; 2012. Available from: https://era.library.ualberta.ca/files/hh63sw19n

University of Ottawa

2. Eason, Matthew. A GFP-Based Sensor to Detect Transiently Expressed Proteins .

Degree: 2020, University of Ottawa

URL: http://hdl.handle.net/10393/40500

► Green fluorescent protein (GFP) fusion tags are commonly used to study protein expression and cellular localization in vivo. But, GFP must undergo an autogenic post-translational… (more)

Subjects/Keywords: Biosensor; Fluorescent Protein; Protein Engineering

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Eason, M. (2020). A GFP-Based Sensor to Detect Transiently Expressed Proteins . (Thesis). University of Ottawa. Retrieved from http://hdl.handle.net/10393/40500

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):

Eason, Matthew. “A GFP-Based Sensor to Detect Transiently Expressed Proteins .” 2020. Thesis, University of Ottawa. Accessed April 11, 2021. http://hdl.handle.net/10393/40500.

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

MLA Handbook (7^th Edition):

Eason, Matthew. “A GFP-Based Sensor to Detect Transiently Expressed Proteins .” 2020. Web. 11 Apr 2021.

Vancouver:

Eason M. A GFP-Based Sensor to Detect Transiently Expressed Proteins . [Internet] [Thesis]. University of Ottawa; 2020. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/10393/40500.

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

Council of Science Editors:

Eason M. A GFP-Based Sensor to Detect Transiently Expressed Proteins . [Thesis]. University of Ottawa; 2020. Available from: http://hdl.handle.net/10393/40500

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

University of Alberta

3. Carlson, Haley J. Development of cpRFP's for use as Ca2+ biosensors.

Degree: PhD, Department of Chemistry, 2013, University of Alberta

URL: https://era.library.ualberta.ca/files/5138jf10b

► The discovery of green fluorescent protein (GFP) from the Aequorea victoria jellyfish revolutionized many fields in the scientific community, including molecular biology, protein engineering, and… (more)

▼ The discovery of green fluorescent protein (GFP) from the Aequorea victoria jellyfish revolutionized many fields in the scientific community, including molecular biology, protein engineering, and neuroscience. The ability to genetically link a fluorescent protein to a protein of interest has allowed scientists to probe the exact structural localization of proteins. Another important application of FPs is their design for use in biosensors, whereby the fluorescence of the protein is intrinsically dependent on a small molecule of interest, such as calcium ion (Ca2+) or a physiological process such as phosphorylation or caspase activity. In single FP-based biosensors of small molecules, the FP must be circular permutated, whereby the original N- and C-termini are linked together and new termini are introduced closer to the chromophore. At the start of the work described in this thesis a lot of work had gone into developing and improving GFP-based Ca2+ biosensors1,2, but there were no reports of a red FP-based biosensor. The work in this thesis describes the engineering of an RFP-based Ca2+ biosensor using a circular permutated RFP, mCherry. The first step in this process was to engineer a cpmCherry variant with termini near the chromophore3. mCherry required a lot of engineering and optimization in order to identify a fluorescent variant with termini near the chromophore. Ultimately, a cpmCherry split at position 145 was found that, when fused to calmodulin (CaM) and M13, showed a response to Ca2+. The initial construct had limited response and was subjected to several rounds of mutagenesis to improve both the brightness and fluorescence response. The final variant CH-GECO3.1 shows a 250% signal increase with Ca2+ and could be imaged successfully in mammalian cells to monitor Ca2+ fluctuations. To further our understanding of this biosensor, site-directed mutagenesis was done to probe the structure-function relationship. After mutagenesis a few residues stood out as key residues that likely played a role in the mechanism of fluorescence increase, such as Gln163 and Glu61 (linker). Other mutations were introduced into the protein to determine whether the excitation and emission wavelengths could be altered, while still retaining function. The final section of this work describes the reconstitution of split green and red Ca2+ biosensors using intein technology. Inteins will spontaneously splice together protein fragments that are genetically linked to them. To take advantage of protein splicing several different Ca2+ biosensors were split into and N-terminal and C-terminal fragments and attached to the N-terminal or C-terminal intein, respectively. These fragments were co-transfected into mammalian HeLa cells and imaged for fluorescence signal and response to Ca2+ fluctuations.

Subjects/Keywords: biosensor; cpRFP; fluorescent protein

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Carlson, H. J. (2013). Development of cpRFP's for use as Ca2+ biosensors. (Doctoral Dissertation). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/5138jf10b

Chicago Manual of Style (16^th Edition):

Carlson, Haley J. “Development of cpRFP's for use as Ca2+ biosensors.” 2013. Doctoral Dissertation, University of Alberta. Accessed April 11, 2021. https://era.library.ualberta.ca/files/5138jf10b.

MLA Handbook (7^th Edition):

Carlson, Haley J. “Development of cpRFP's for use as Ca2+ biosensors.” 2013. Web. 11 Apr 2021.

Vancouver:

Carlson HJ. Development of cpRFP's for use as Ca2+ biosensors. [Internet] [Doctoral dissertation]. University of Alberta; 2013. [cited 2021 Apr 11]. Available from: https://era.library.ualberta.ca/files/5138jf10b.

Council of Science Editors:

Carlson HJ. Development of cpRFP's for use as Ca2+ biosensors. [Doctoral Dissertation]. University of Alberta; 2013. Available from: https://era.library.ualberta.ca/files/5138jf10b

King Abdullah University of Science and Technology

4. Fischer, Johannes. A Pathway to Artificial Metalloenzymes.

Degree: Biological and Environmental Sciences and Engineering (BESE) Division, 2015, King Abdullah University of Science and Technology

URL: http://hdl.handle.net/10754/583809

► The advancement of catalytic systems and the application thereof has proven to be the key to overcome traditional limitations of industrial-scale synthetic processes. Converging organometallic… (more)

▼ The advancement of catalytic systems and the application thereof has proven to be the key to overcome traditional limitations of industrial-scale synthetic processes. Converging organometallic and biocatalytic principles lead to the development of Artificial Metalloenzymes (ArMs) that comprise a synthetic metal catalyst embedded in a protein scaffold, thereby combining the reactivity of the former with the versatility of the latter. This synergistic approach introduces rationally designed building blocks for the catalytic site and the host protein to assemble enzyme-like structures that follow regio-, chemo-, enantio- and substrate-selective principles. Yet, the identification of suitable protein scaffolds has thus far been challenging. Herein we report a rationally optimized fluorescent protein host, mTFP*, that was engineered to have no intrinsic metal binding capability and, owing to its robust nature, can act as scaffold for the design of novel ArMs. We demonstrate the potential of site-specific modifications within the protein host, use protein X-Ray analysis to validate the respective scaffolds and show how artificial mutant binding sites can be introduced. Transition metal Förster Resonance Energy transfer (tmFRET) methodologies help to evaluate micromolar dissociation constants and reveal structural rearrangements upon coordination of the metal centers. In conjunction with molecular insights from X-Ray crystallographic structure determination, dynamics of the binding pocket can be inferred. The versatile subset of different binding motifs paired with transition metal catalysts create artificial metalloenzymes that provide reactivities which otherwise do not exist in nature. As a proof of concept, Diels-Alder cycloadditions highlight the potential of the present mTFP* based catalysts by stereoselectively converting azachalcone and cyclopentadiene substrates. Screens indicate an enantiomeric excess of up to 60% and provide insights into the electronic and geometric constitution of the first coordination spheres binding the catalysts. We further apply two general principles to optimize selective conversions of the generated ArMs. 1) Utilizing site-specific mutagenesis, increased hydrophobicity is introduced to the second coordination sphere. 2) In-vitro post-expressional modification utilizing N-hydroxysuccinimide esters is anticipated to introduce a sterically more demanding second coordination sphere that influences substrate entry by favoring a particular stereoisomer. The latter approach however also enhances the host proteins robustness under processing conditions. The presented study investigates a novel approach to create artificial metalloenzymes based on non-enzymatic precursor proteins. It illustrates means of modification and functionalization. Further guidance to overcome the general problem of insufficient stereoselectivity and stability is also presented. In view of the insights gained we see the importance of further mutagenic studies, i.e. through means of guided… Advisors/Committee Members: Eppinger, Jörg (advisor), Arold, Stefan T. (committee member), Groll, Michael (committee member), Hamdan, Samir (committee member), Uwe, Bunz (committee member).

Subjects/Keywords: Metalloenzymes; fluorescent protein; Diels-Alder

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Fischer, J. (2015). A Pathway to Artificial Metalloenzymes. (Thesis). King Abdullah University of Science and Technology. Retrieved from http://hdl.handle.net/10754/583809

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):

Fischer, Johannes. “A Pathway to Artificial Metalloenzymes.” 2015. Thesis, King Abdullah University of Science and Technology. Accessed April 11, 2021. http://hdl.handle.net/10754/583809.

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

MLA Handbook (7^th Edition):

Fischer, Johannes. “A Pathway to Artificial Metalloenzymes.” 2015. Web. 11 Apr 2021.

Vancouver:

Fischer J. A Pathway to Artificial Metalloenzymes. [Internet] [Thesis]. King Abdullah University of Science and Technology; 2015. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/10754/583809.

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

Council of Science Editors:

Fischer J. A Pathway to Artificial Metalloenzymes. [Thesis]. King Abdullah University of Science and Technology; 2015. Available from: http://hdl.handle.net/10754/583809

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

University of Rochester

5. Dean, Kimberly Marie. Selection, Identification, and Characterization of Codon Pairs that Inhibit Translation and the Development of the RNA-ID Method to Measure the Effects of Cis-Regulatory Elements.

Degree: PhD, 2013, University of Rochester

URL: http://hdl.handle.net/1802/27301

► It has been known for over 30 years that synonymous codon choice regulates translation, but the characteristics of codons that inhibit translation, and the factors… (more)

▼ It has been known for over 30 years that synonymous codon choice regulates translation, but the characteristics of codons that inhibit translation, and the factors that restrict their function, are unknown. In a systematic screen of 59 of 61 sense codons, our lab identified the arginine CGA codon as strongly inhibitory, and then determined that CGA inhibition was primarily due to A·I wobble decoding. Furthermore, I found that adjacent CGA codons are synergistically inhibitory relative to separated CGA codons, which presents the possibility that combinations of non-identical codons may regulate translation in ways that are not predictable from their individual codon behavior. To search for inhibitory codon combinations among the 3,721 codon pairs and 226,981 codon triplets, I developed the RNA-ID method to screen for cis-regulatory elements. RNA-ID utilizes a fluorescence-based, integrated reporter and coupled to flow cytometry to evaluate effects of sequences on expression in Saccharomyces cerevisiae. This method is useful because insertion of test sequences, using ligation independent cloning, is simple, expression is detectable over a 250-fold range, measurements are quantitative and dose-dependent, separation of specific populations is nearly complete, and results from a single sequence are reproducible. To find inhibitory codon combinations, a library of sequences, consisting of random 9 nucleotide inserts, was evaluated using RNA-ID. Only a small fraction of yeast (<0.2%) exhibited GFP/RFP at 6-10% the maximal level, indicating that very few sequences inhibit expression to this degree. Ninety-two strains from this group were studied. Five novel sequences that inhibit translation where identified based on suppression of the expression defect by appropriate tRNAs. The inhibitory sequences all contain at least one codon decoded by wobble, and in each case, the suppression of the inhibitory effect requires expression of a mutated, exact base-pairing tRNA. Thus, I conclude that wobble decoding is likely to be a critical factor in their inhibition of translation. In two cases, inhibition depends upon a pair of adjacent codons and upon the order of these codons, thus indicating that the intact codon pair is the inhibitory unit, consistent with the idea that they act within a single round of translation.

Subjects/Keywords: Yeast; Genetic Code; Fluorescence, Green Fluorescent Protein; Red Fluorescent Protein

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Dean, K. M. (2013). Selection, Identification, and Characterization of Codon Pairs that Inhibit Translation and the Development of the RNA-ID Method to Measure the Effects of Cis-Regulatory Elements. (Doctoral Dissertation). University of Rochester. Retrieved from http://hdl.handle.net/1802/27301

Chicago Manual of Style (16^th Edition):

Dean, Kimberly Marie. “Selection, Identification, and Characterization of Codon Pairs that Inhibit Translation and the Development of the RNA-ID Method to Measure the Effects of Cis-Regulatory Elements.” 2013. Doctoral Dissertation, University of Rochester. Accessed April 11, 2021. http://hdl.handle.net/1802/27301.

MLA Handbook (7^th Edition):

Dean, Kimberly Marie. “Selection, Identification, and Characterization of Codon Pairs that Inhibit Translation and the Development of the RNA-ID Method to Measure the Effects of Cis-Regulatory Elements.” 2013. Web. 11 Apr 2021.

Vancouver:

Dean KM. Selection, Identification, and Characterization of Codon Pairs that Inhibit Translation and the Development of the RNA-ID Method to Measure the Effects of Cis-Regulatory Elements. [Internet] [Doctoral dissertation]. University of Rochester; 2013. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/1802/27301.

Council of Science Editors:

Dean KM. Selection, Identification, and Characterization of Codon Pairs that Inhibit Translation and the Development of the RNA-ID Method to Measure the Effects of Cis-Regulatory Elements. [Doctoral Dissertation]. University of Rochester; 2013. Available from: http://hdl.handle.net/1802/27301

Michigan State University

6. Assar, Zahra. Elucidation of iLBP family folding pathway and study of reengineering them as fluorescent protein tags via structural analysis.

Degree: 2018, Michigan State University

URL: http://etd.lib.msu.edu/islandora/object/etd:6825

►

"The intracellular lipid binding proteins (iLBP) family are found in the cells of mammals, birds, fish, amphibians and reptiles. They function to shuttle large insoluble… (more)

▼

"The intracellular lipid binding proteins (iLBP) family are found in the cells of mammals, birds, fish, amphibians and reptiles. They function to shuttle large insoluble hydrophobic molecules, including retinal, various long chain fatty acids and etc. throughout the cell around the cytosol and nucleus. The combination of their small size and relatively large binding pocket make them suitable templates in a variety of protein design applications, including the study of an innovative class of fluorescent proteins. To pursue our goals, we used human Cellular Retinol Binding Protein II (hCRBPII). We were the first group to achieve the structure of an all-transretinal and the first bonafide structure of retinol-bound hCRBPII. In the course of these studies, we have discovered hCRBPII is surprisingly capable of folding as domain swapped dimer (DS), with single mutations able to shift the folding product from monomer to dimer. Structural analysis of both wild type and multiple mutant DS dimers led us to remarkable hypotheses regarding mechanism of this phenomenon, which is different from the previous studies on this family. We proposed that the N-terminal and C-terminal halves of hCRBPII are capable of at least partially folding independently, to form "open monomer." The dimer/monomer ratio depends on the relative rates of dimerization of the open monomers, versus closing of the two halves together to form the "closed monomer". In addition, by comparing structures of holo hCRBPII DS dimer variants, we identified an extremely large change in the relative orientation of the two domains upon ligand binding in dimers. This suggests the possibility that iLBP domain swapped dimers could be allosterically regulated forms of these proteins, at least in some cases. Fatty acid binding protein 5 (FABP5), another member of iLBPs, has also been reported to forms a very similar DS dimer, which makes it likely that other family members could also form DS dimers, and have physiological importance for some members of the family. As mentioned, a new class of fluorogenic proteins was created by binding fluorophore aldehydes in the binding pocket of hCRBPII via protonated Schiff base (PSB) formation. In this new system, emission of the designed solvatochromic fluorophore is flexible based on the polarity of the environment; therefore multicolor probes can be developed. More importantly, absorption/emission wavelengths can be tuned therefor; nonspecific labeling and background fluorescence can be reduced. By now, the absorption maxima are tuned from 501nm to 705nm and emission maxima from 613 nm to 744 nm. Covering both the red and far-red fluorescence wavelength regimes." – Pages ii-iii.

Online resource;

Advisors/Committee Members: Geiger, James H., Borhan, Babak, Blanchard, Gary, Henry, William.

Subjects/Keywords: Lipoproteins; Protein engineering; Protein folding; Fluorescent polymers; Fluorescent probes; Chemistry; Biochemistry

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Assar, Z. (2018). Elucidation of iLBP family folding pathway and study of reengineering them as fluorescent protein tags via structural analysis. (Thesis). Michigan State University. Retrieved from http://etd.lib.msu.edu/islandora/object/etd:6825

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):

Assar, Zahra. “Elucidation of iLBP family folding pathway and study of reengineering them as fluorescent protein tags via structural analysis.” 2018. Thesis, Michigan State University. Accessed April 11, 2021. http://etd.lib.msu.edu/islandora/object/etd:6825.

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

MLA Handbook (7^th Edition):

Assar, Zahra. “Elucidation of iLBP family folding pathway and study of reengineering them as fluorescent protein tags via structural analysis.” 2018. Web. 11 Apr 2021.

Vancouver:

Assar Z. Elucidation of iLBP family folding pathway and study of reengineering them as fluorescent protein tags via structural analysis. [Internet] [Thesis]. Michigan State University; 2018. [cited 2021 Apr 11]. Available from: http://etd.lib.msu.edu/islandora/object/etd:6825.

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

Council of Science Editors:

Assar Z. Elucidation of iLBP family folding pathway and study of reengineering them as fluorescent protein tags via structural analysis. [Thesis]. Michigan State University; 2018. Available from: http://etd.lib.msu.edu/islandora/object/etd:6825

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

Hong Kong University of Science and Technology

7. Jiang, Meijuan CHEM. Development of novel AIEgens based on benzylidene-methyloxazolones and isoquinolinium salts and exploration of their biological applications.

Degree: 2017, Hong Kong University of Science and Technology

URL: http://repository.ust.hk/ir/Record/1783.1-95153 ; https://doi.org/10.14711/thesis-991012564469003412 ; http://repository.ust.hk/ir/bitstream/1783.1-95153/1/th_redirect.html

► To tackle world-challenging problems, luminescent materials have played a vital role in the promotion of scientific discoveries and technological innovations. Practically, traditional luminogens often face… (more)

▼ To tackle world-challenging problems, luminescent materials have played a vital role in the promotion of scientific discoveries and technological innovations. Practically, traditional luminogens often face a problem of aggregation-caused quenching (ACQ), resulting a reduction or diminishment of luminescence and thus limiting their practical applications. Circumventing the notorious ACQ effect, luminogens with aggregation-induced emission characteristics (AIEgens) have become intriguing with a rapid expansion in applications ranging from optoelectronics, chemo/biosensing to bioimaging. However, to tailor for specific high-tech applications and deepen the mechanistic understanding of AIEgens, there is a high demand on the developments of novel AIEgens with easy preparation and functionalization. In this thesis, two series of AIEgens were developed. Their working mechanism and various applications, especially biological applications, were investigated. Based on green fluorescence protein chromophore analogues benzylidene-oxazolone, a series of AIEgens were synthesized with solid quantum yield of up to 50% and emission wavelength of up to 635 nm. Their working mechanism were carefully deciphered. Further, through minor modifications, a two-photon AIE bioprobe was developed for specific lipid droplet imaging in cells and tissues. Also, we prepared a series of isoquinolinium salts-based AIEgens with high structural stability via a simple one-pot reaction. With the merits of visible light excitation, large Stokes shift, high quantum yield, tunable color and sufficient two-photon absorption of near-infrared light, we found their various applications in mechanochromic rewritable paper, mitochondrial targeting cell imaging and bacterial imaging. Interestingly, these molecules exhibited high viscosity-sensitivity and were further utilized as sensors for extracellular and intracellular microviscosity sensing. Among them, a simple AIEgen was developed for image-guided two-photon excited photodymanic therapy for precise cancer treatment.

Subjects/Keywords: Electroluminescent devices ; Photoemission ; Aggregation (Chemistry) ; Green fluorescent protein ; Fluorescent polymers ; Isoquinoline

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Jiang, M. C. (2017). Development of novel AIEgens based on benzylidene-methyloxazolones and isoquinolinium salts and exploration of their biological applications. (Thesis). Hong Kong University of Science and Technology. Retrieved from http://repository.ust.hk/ir/Record/1783.1-95153 ; https://doi.org/10.14711/thesis-991012564469003412 ; http://repository.ust.hk/ir/bitstream/1783.1-95153/1/th_redirect.html

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):

Jiang, Meijuan CHEM. “Development of novel AIEgens based on benzylidene-methyloxazolones and isoquinolinium salts and exploration of their biological applications.” 2017. Thesis, Hong Kong University of Science and Technology. Accessed April 11, 2021. http://repository.ust.hk/ir/Record/1783.1-95153 ; https://doi.org/10.14711/thesis-991012564469003412 ; http://repository.ust.hk/ir/bitstream/1783.1-95153/1/th_redirect.html.

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

MLA Handbook (7^th Edition):

Jiang, Meijuan CHEM. “Development of novel AIEgens based on benzylidene-methyloxazolones and isoquinolinium salts and exploration of their biological applications.” 2017. Web. 11 Apr 2021.

Vancouver:

Jiang MC. Development of novel AIEgens based on benzylidene-methyloxazolones and isoquinolinium salts and exploration of their biological applications. [Internet] [Thesis]. Hong Kong University of Science and Technology; 2017. [cited 2021 Apr 11]. Available from: http://repository.ust.hk/ir/Record/1783.1-95153 ; https://doi.org/10.14711/thesis-991012564469003412 ; http://repository.ust.hk/ir/bitstream/1783.1-95153/1/th_redirect.html.

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

Council of Science Editors:

Jiang MC. Development of novel AIEgens based on benzylidene-methyloxazolones and isoquinolinium salts and exploration of their biological applications. [Thesis]. Hong Kong University of Science and Technology; 2017. Available from: http://repository.ust.hk/ir/Record/1783.1-95153 ; https://doi.org/10.14711/thesis-991012564469003412 ; http://repository.ust.hk/ir/bitstream/1783.1-95153/1/th_redirect.html

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

Michigan State University

8. Santos, Elizabeth Marie. Development of fluorescent protein tags for live-cell imaging.

Degree: 2017, Michigan State University

URL: http://etd.lib.msu.edu/islandora/object/etd:6843

►

"Our primary goal is to develop fluorescent proteins that span the entire visible spectra, to be used when conventional fluorescent proteins are inadequate. In our… (more)

▼

"Our primary goal is to develop fluorescent proteins that span the entire visible spectra, to be used when conventional fluorescent proteins are inadequate. In our system, fluorescence is activated upon coupling of the protein and ligand, such that temporal control can be achieved, whereas intrinsically fluorescent proteins are constitutively on. Additionally, our system does not require oxygen and can therefore find potential uses in obligate anaerobes. Our lab has demonstrated the ability to effectively control the absorption profile of conjugated polyenes. The initial aim of this project was to regulate the emissive properties of bound fluorophores with the same degree of control. This was achieved by the coupling of the solvatochromic fluorophore ThioFluor to hCRBPII mutants. ThioFluor yielded mutants with absorption maxima varying from 501 nm to 705 nm and emission maxima from 613 nm to 744 nm. This is equivalent to regulation over 204 nm in absorption and 131 nm in emission, covering both the red and far-red fluorescence wavelength regimes. Furthermore, we have shown its utility in live-cell imaging in whole cells, and with targeting to the nucleus and extranuclear space; fortuitously, negligible background fluorescence is apparent. In the course of optimizing binding of ThioFluor to hCRBPII, we discovered that it was observed that not only is the protonated Schiff base (PSB) fluorescent, but the Schiff base (SB) is as well. However, while PSB emission wavelength could be altered over 204 nm, SB emission remains nearly constant at 500 nm. Interestingly, select mutants displayed a far-red emission upon SB irradiation, similar to that obtained upon irradiation of the PSB, presumably through protonation in the excited state. This serendipitous discovery leads to more than a 200 nm Stokes shift with high quantum yield (> 60%). One such hCRBPII/ThioFluor complex displays ideal spectroscopic properties including fast iminium formation with a half-life of 1.7 min, low pKa of 5.3 (rendering almost complete SB formation at physiological pH), high quantum yield (0.51) and large Stokes shift (208 nm). This fluorescent protein was successfully used to visualize whole cell fluorescence, as well as targeting to the nucleus. The last major endeavor was to develop a protein-based pH sensor, with the ability to report pH values with high accuracy. We have previously reported an absorptive system that was capable of ratiometric sensing of pH. However, we have now developed a single protein fluorescent ratiometric pH sensor, based on the titration of an acidic residue near the iminium. Standard curves were generated based on the ratio of emissions at two excitation wavelengths, allowing for concentration independent sensing of pH. We have been able to demonstrate its applicability as an in vivo fluorescent pH probe, obtaining a pH value of 6.7 when hCRBPII is targeted to the nucleus of HeLa cells." – Pages ii-iii.

Online resource;

Advisors/Committee Members: Borhan, Babak, Jackson, James, Geiger, James, Huang, Xuefei.

Subjects/Keywords: Fluorescent polymers – Research; Fluorescent probes; Protein engineering; Organic chemistry; Biochemistry

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Santos, E. M. (2017). Development of fluorescent protein tags for live-cell imaging. (Thesis). Michigan State University. Retrieved from http://etd.lib.msu.edu/islandora/object/etd:6843

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):

Santos, Elizabeth Marie. “Development of fluorescent protein tags for live-cell imaging.” 2017. Thesis, Michigan State University. Accessed April 11, 2021. http://etd.lib.msu.edu/islandora/object/etd:6843.

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

MLA Handbook (7^th Edition):

Santos, Elizabeth Marie. “Development of fluorescent protein tags for live-cell imaging.” 2017. Web. 11 Apr 2021.

Vancouver:

Santos EM. Development of fluorescent protein tags for live-cell imaging. [Internet] [Thesis]. Michigan State University; 2017. [cited 2021 Apr 11]. Available from: http://etd.lib.msu.edu/islandora/object/etd:6843.

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

Council of Science Editors:

Santos EM. Development of fluorescent protein tags for live-cell imaging. [Thesis]. Michigan State University; 2017. Available from: http://etd.lib.msu.edu/islandora/object/etd:6843

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

University of Texas – Austin

9. Hunt, Marguerite E. Exploring the emerging properties of novel GFP-like fluorescent proteins.

Degree: MA, Cell and Molecular Biology, 2013, University of Texas – Austin

URL: http://hdl.handle.net/2152/22311

► In 2008 the Nobel Prize in Chemistry was awarded to the scientists who revolutionized biomedical technology by isolating, characterizing, and pioneering the use of a… (more)

▼ In 2008 the Nobel Prize in Chemistry was awarded to the scientists who revolutionized biomedical technology by isolating, characterizing, and pioneering the use of a green fluorescent protein (GFP) from a humble hydrozoan jellyfish. Now numbering in the hundreds of colors and applications, fluorescent protein (FP) tools have facilitated the explosion of biological knowledge elucidated by a technology that can label DNA or RNA, track protein expression, and identify protein interactions. The development of the large variety of FP biotechnology available today has been due to the need for expanded color palettes and applications, and more efficient functionality. Yet, as our understanding of the biochemical and spectral characteristics of these genetically-encoded, self-assembling proteins has expanded, our comprehension of the biological function of FPs in the host organisms has remained inadequate. While the need for novel FP laboratory applications still continues, the new focus in the field of fluorescent proteins is moving to also characterize their biological functions. In this research compilation, the identification of three groups of new fluorescent proteins from marine copepods and hydrozoans has provided a collection of eleven FPs exhibiting previously uncharacterized colors, and biochemical and structural features. The green FPs from copepods are the brightest wild-type FPs identified and support the hypothesized biological function of fluorescence as counter-shading in the marine environment where these animals live. The FPs from the siphonophore and anthoathecate jelly, both hydrozoan animals, are comprised of tandemly expressed fluorescent protein units, a solution to the oligomeric structure common to most FPs that suggests a novel structure-function relationship. The fluorescent proteins from Obelia reveal a novel hydrozoan cyan FP, previously uncharacterized higher-order structural complexes, and have initiated the work to describe the biological function of these proteins as potential regenerators of their internal bioluminescent light sources. All eleven fluorescent proteins may also be adapted for FP technology. Advisors/Committee Members: Matz, Mikhail V. (advisor).

Subjects/Keywords: Fluorescent protein; Biotechnology; Fluorescent spectra; Genetically encoded proteins

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Hunt, M. E. (2013). Exploring the emerging properties of novel GFP-like fluorescent proteins. (Masters Thesis). University of Texas – Austin. Retrieved from http://hdl.handle.net/2152/22311

Chicago Manual of Style (16^th Edition):

Hunt, Marguerite E. “Exploring the emerging properties of novel GFP-like fluorescent proteins.” 2013. Masters Thesis, University of Texas – Austin. Accessed April 11, 2021. http://hdl.handle.net/2152/22311.

MLA Handbook (7^th Edition):

Hunt, Marguerite E. “Exploring the emerging properties of novel GFP-like fluorescent proteins.” 2013. Web. 11 Apr 2021.

Vancouver:

Hunt ME. Exploring the emerging properties of novel GFP-like fluorescent proteins. [Internet] [Masters thesis]. University of Texas – Austin; 2013. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/2152/22311.

Council of Science Editors:

Hunt ME. Exploring the emerging properties of novel GFP-like fluorescent proteins. [Masters Thesis]. University of Texas – Austin; 2013. Available from: http://hdl.handle.net/2152/22311

University of Alberta

10. Shen, Yi. Engineering of red fluorescent proteins and red fluorescent protein-based pH biosensors.

Degree: PhD, Department of Chemistry, 2014, University of Alberta

URL: https://era.library.ualberta.ca/files/c9z902z87p

► Fluorescent proteins (FP) and FP-based biosensors have become essential tools for cell biology and neuroscience research. This thesis describes efforts to engineer new FPs with… (more)

▼ Fluorescent proteins (FP) and FP-based biosensors have become essential tools for cell biology and neuroscience research. This thesis describes efforts to engineer new FPs with red emission and novel genetically encoded biosensors based on red FPs. Directed evolution and semi-rational design are the main techniques used to develop novel and improved red FP and red FP-based biosensors. First, A new pH-sensitive red fluorescent protein based on mApple, termed as “pHuji”, has been developed with high pH sensitivity, exhibiting over 20-fold fluorescence intensity change between pH 5.5 and pH 7.5. In live cell imaging, cell surface-displayed pHuji demonstrated high pH sensitivity when exposed to buffers with defined pH values. Collaborators have used pHuji for successful visualization of pH changes during exocytosis and endocytosis. Following the engineering of intensiometric red pH sensors, we next turned our attention to ratiometric red pH sensors. Through a process of semi-rational design and directed evolution, the red FP mApple was successfully engineered into a series of dual-excitation, ratiometric pH sensors. These new red-shifted ratiometric pH sensors, termed pHlorina, exhibit large ratio change (over 70-fold) for pH changes from 5.0 to 7.5. A series of long Stokes shift variants of mApple (λex = 450 nm and λem = 610 nm) were also developed and characterized. A photochromic and thermochromic red FP was serendipitously discovered during the process of engineering the long Stokes shift red FP. This protein, which we designated as switchable hypersensitive red FP (shyRFP), was characterized in terms of light, temperature, and pH dependence. A colour switching mechanism that involves protonation coupled E-Z isomerization of the protein chromophore was proposed. The monomeric RFP mCherry is widely used for live cell fluorescence imaging experiments. Using semi-rational design and random mutagenesis, two new mCherry variants were developed: Long Stokes Shift mCherry (LSSmCherry; λex = 460 nm and λem = 610 nm) and Red-Shifted mCherry (RDSmCherry; λex = 600 nm and λem = 630 nm). These two proteins have distinctively different fluorescence excitation and emission profiles from their predecessor mCherry2. These new additions to the FP toolbox provide templates for the engineering of new colour variants and fluorescent sensors with red emission. Finally, a filter paper-based screening method has been developed to screen for calcium ion (Ca2+) sensitive RFP variants expressed in Escherichia coli colonies. These low-affinity Ca2+ sensors were semi-rationally designed by altering protein barrel residues near the chromophore to form a Ca2+ binding pocket. By combining high-throughput screening and rational design, a number of Ca2+ sensitive variants were successfully identified.

Subjects/Keywords: red fluorescent proteins; protein engineering; biosensors

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Shen, Y. (2014). Engineering of red fluorescent proteins and red fluorescent protein-based pH biosensors. (Doctoral Dissertation). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/c9z902z87p

Chicago Manual of Style (16^th Edition):

Shen, Yi. “Engineering of red fluorescent proteins and red fluorescent protein-based pH biosensors.” 2014. Doctoral Dissertation, University of Alberta. Accessed April 11, 2021. https://era.library.ualberta.ca/files/c9z902z87p.

MLA Handbook (7^th Edition):

Shen, Yi. “Engineering of red fluorescent proteins and red fluorescent protein-based pH biosensors.” 2014. Web. 11 Apr 2021.

Vancouver:

Shen Y. Engineering of red fluorescent proteins and red fluorescent protein-based pH biosensors. [Internet] [Doctoral dissertation]. University of Alberta; 2014. [cited 2021 Apr 11]. Available from: https://era.library.ualberta.ca/files/c9z902z87p.

Council of Science Editors:

Shen Y. Engineering of red fluorescent proteins and red fluorescent protein-based pH biosensors. [Doctoral Dissertation]. University of Alberta; 2014. Available from: https://era.library.ualberta.ca/files/c9z902z87p

Northeastern University

11. Salna, Bridget I. Proton transport in proteins and the role of quantum tunneling.

Degree: PhD, Department of Physics, 2017, Northeastern University

URL: http://hdl.handle.net/2047/D20248978

► Proton transport is ubiquitous in biological systems and has been studied extensively, both with experimental and theoretical methods. A better understanding of this phenomenon is… (more)

▼ Proton transport is ubiquitous in biological systems and has been studied extensively, both with experimental and theoretical methods. A better understanding of this phenomenon is sought because it underpins many fundamental mechanisms that sustain life, such as cellular respiration, photosynthesis, and drug metabolism. Many of these processes involve proton wires within proteins, in which protons are transported for use in biochemical reactions. This work examines the contribution of quantum tunneling to proton transport and proposes a primary role for tunneling in regulating this process.; This work discusses and improves upon the current theoretical models of proton tunneling in proteins and investigates the implications of considering tunneling to be the dominant transport channel. By including deep quantum tunneling under high potential barriers as a viable proton transport pathway, ionization-resistant residues such as serine and threonine can be considered as active constituents of proton wires. These residues have previously been found along many proton wires in proteins, but are seldom identified as transport elements. One notable exception is the green fluorescent protein (GFP), in which serine has been established as an active element of the well-characterized internal proton wire. This makes GFP an important model system to test the role of quantum tunneling in proton transport within proteins. This is one of the main projects discussed in this work, in which the biologically relevant ground state process is considered in depth. The rate-limiting step at room temperature is assigned to deep tunneling from the serine hydroxyl with a rate that is orders of magnitude faster than the classical pathway. This suggests how high pK_a residues can act to stabilize and regulate proton flow along proton wires in proteins.; The role of quantum tunneling in enzymatic systems is also explored, in which proton coupled electron transfer (PCET) is often the critical process. The PCET in soybean lipoxygenase-1 (SLO), as well as its double mutant, is analyzed using a modified model of donor-acceptor atom interaction. This includes effects of electronic repulsion, external protein forces, and charge/bond polarization. Distinct mechanisms are proposed for the two species of SLO, in which the wild type forms an activated state with the substrate positioned close to the active site, while the double mutant is constrained to longer distances and transfer is only possible due to its increased flexibility.

Subjects/Keywords: green fluorescent protein; proton transport; tunneling

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Salna, B. I. (2017). Proton transport in proteins and the role of quantum tunneling. (Doctoral Dissertation). Northeastern University. Retrieved from http://hdl.handle.net/2047/D20248978

Chicago Manual of Style (16^th Edition):

Salna, Bridget I. “Proton transport in proteins and the role of quantum tunneling.” 2017. Doctoral Dissertation, Northeastern University. Accessed April 11, 2021. http://hdl.handle.net/2047/D20248978.

MLA Handbook (7^th Edition):

Salna, Bridget I. “Proton transport in proteins and the role of quantum tunneling.” 2017. Web. 11 Apr 2021.

Vancouver:

Salna BI. Proton transport in proteins and the role of quantum tunneling. [Internet] [Doctoral dissertation]. Northeastern University; 2017. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/2047/D20248978.

Council of Science Editors:

Salna BI. Proton transport in proteins and the role of quantum tunneling. [Doctoral Dissertation]. Northeastern University; 2017. Available from: http://hdl.handle.net/2047/D20248978

Brandeis University

12. Yanding, Zhao. Fluorescent labeled TAL protein as a tool to investigate single-molecule transcription.

Degree: 2015, Brandeis University

URL: http://hdl.handle.net/10192/31114

► TALEs are sequence specific transcription activator found in phytopathogenic bacteria. Interestingly, their specificity is defined by triplet amino acid repeat sequences, which can recognize individual… (more)

Subjects/Keywords: Fluorescent; TAL protein; single-molecule transcription

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Yanding, Z. (2015). Fluorescent labeled TAL protein as a tool to investigate single-molecule transcription. (Thesis). Brandeis University. Retrieved from http://hdl.handle.net/10192/31114

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):

Yanding, Zhao. “Fluorescent labeled TAL protein as a tool to investigate single-molecule transcription.” 2015. Thesis, Brandeis University. Accessed April 11, 2021. http://hdl.handle.net/10192/31114.

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

MLA Handbook (7^th Edition):

Yanding, Zhao. “Fluorescent labeled TAL protein as a tool to investigate single-molecule transcription.” 2015. Web. 11 Apr 2021.

Vancouver:

Yanding Z. Fluorescent labeled TAL protein as a tool to investigate single-molecule transcription. [Internet] [Thesis]. Brandeis University; 2015. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/10192/31114.

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

Council of Science Editors:

Yanding Z. Fluorescent labeled TAL protein as a tool to investigate single-molecule transcription. [Thesis]. Brandeis University; 2015. Available from: http://hdl.handle.net/10192/31114

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

University of Kansas

13. Lei, Ming. Using Lysine-Reactive Fluorescent Dye for Surface Characterization of a Monoclonal Antibody.

Degree: MA, Pharmaceutical Chemistry, 2014, University of Kansas

URL: http://hdl.handle.net/1808/21645

► The last decade has witnessed a rapid growth in the development of protein pharmaceuticals for diagnostic and therapeutic purposes. The biopharmaceutical industry increasingly demands thorough… (more)

Subjects/Keywords: Chemistry; Fluorescent; labeling; mass spectrometry; protein structure

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Lei, M. (2014). Using Lysine-Reactive Fluorescent Dye for Surface Characterization of a Monoclonal Antibody. (Masters Thesis). University of Kansas. Retrieved from http://hdl.handle.net/1808/21645

Chicago Manual of Style (16^th Edition):

Lei, Ming. “Using Lysine-Reactive Fluorescent Dye for Surface Characterization of a Monoclonal Antibody.” 2014. Masters Thesis, University of Kansas. Accessed April 11, 2021. http://hdl.handle.net/1808/21645.

MLA Handbook (7^th Edition):

Lei, Ming. “Using Lysine-Reactive Fluorescent Dye for Surface Characterization of a Monoclonal Antibody.” 2014. Web. 11 Apr 2021.

Vancouver:

Lei M. Using Lysine-Reactive Fluorescent Dye for Surface Characterization of a Monoclonal Antibody. [Internet] [Masters thesis]. University of Kansas; 2014. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/1808/21645.

Council of Science Editors:

Lei M. Using Lysine-Reactive Fluorescent Dye for Surface Characterization of a Monoclonal Antibody. [Masters Thesis]. University of Kansas; 2014. Available from: http://hdl.handle.net/1808/21645

University of Kansas

14. Egan, Chet. Engineering GFP-Family Member Protiens to Achieve Novel Functions: A Class of Proteins Limited Only by the Imagination.

Degree: MA, Molecular Biosciences, 2015, University of Kansas

URL: http://hdl.handle.net/1808/19178

► The Green Fluorescent Protein (GFP) has single handedly revolutionized microscopy and molecular biology. A simple search for GFP in the NCBI Pubmed database yields over… (more)

▼ The Green Fluorescent Protein (GFP) has single handedly revolutionized microscopy and molecular biology. A simple search for GFP in the NCBI Pubmed database yields over 29,900 unique publications as of April 2015 demonstrating the importance and influence GFP has had on the scientific community. It is because of this impact that the 2008 Nobel Prize in Chemistry was awarded jointly to Osamu Shimomura, Martin Chalfie, and Roger Y. Tsien for their ground breaking work on the discovery and characterization of GFP family proteins. GFP has come a long way since it was first isolated from Aquorea Victoria, a bioluminescent jellyfish native to the Pacific Northwest. In the past 2 decades since it was first cloned countless variants have been engineered. There are now literally hundreds of variants of GFP family proteins with more continuously being engineered. The choice of emission colors spans the entire visible spectrum and even beyond with some reaching into the infrared wavelengths. It would be a daunting task to attempt to cover all the amazing achievements in engineering of GFP family proteins and their diverse applications. Researchers are continuously improving on the constructs available, increasing their efficiency, sensitivity, and stability. GFP family proteins have been adapted to be everything from reporters of gene expression to labels for entire organelles or even entire tissues. GFP can be used to visualize protein binding and transient protein-protein interactions. GFP can also have its fluorescence put under the direct control of a small molecule ligand paving the way for switchable and biosensor GFPs. The more one reads about GFP the more it becomes clear that the applications of this amazing protein family are still expanding at a very rapid pace. Here, I will focus on a few of what I consider the most interesting, ambitious, and novel engineering applications of the GFP family of proteins. Advisors/Committee Members: Karanicolas, John (advisor), Benedict, Steve (cmtemember), Richter, Mark (cmtemember).

Subjects/Keywords: Molecular biology; GFP; Green Fluorescent Protein

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Egan, C. (2015). Engineering GFP-Family Member Protiens to Achieve Novel Functions: A Class of Proteins Limited Only by the Imagination. (Masters Thesis). University of Kansas. Retrieved from http://hdl.handle.net/1808/19178

Chicago Manual of Style (16^th Edition):

Egan, Chet. “Engineering GFP-Family Member Protiens to Achieve Novel Functions: A Class of Proteins Limited Only by the Imagination.” 2015. Masters Thesis, University of Kansas. Accessed April 11, 2021. http://hdl.handle.net/1808/19178.

MLA Handbook (7^th Edition):

Egan, Chet. “Engineering GFP-Family Member Protiens to Achieve Novel Functions: A Class of Proteins Limited Only by the Imagination.” 2015. Web. 11 Apr 2021.

Vancouver:

Egan C. Engineering GFP-Family Member Protiens to Achieve Novel Functions: A Class of Proteins Limited Only by the Imagination. [Internet] [Masters thesis]. University of Kansas; 2015. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/1808/19178.

Council of Science Editors:

Egan C. Engineering GFP-Family Member Protiens to Achieve Novel Functions: A Class of Proteins Limited Only by the Imagination. [Masters Thesis]. University of Kansas; 2015. Available from: http://hdl.handle.net/1808/19178

University of Sydney

15. Morin-Adeline, Victoria. Molecular and Biological Investigations of Tritrichomonas foetus from Cattle, Domestic Cats and Pigs .

Degree: 2016, University of Sydney

URL: http://hdl.handle.net/2123/15168

► Few organisms have the potential to shed light on aspects of parasitism as much as Tritrichomonas foetus. While it is a commensal organism in pigs,… (more)

Subjects/Keywords: Trichomonad; Fluorescent protein; parasite; Transcriptome; cattle; cats

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Morin-Adeline, V. (2016). Molecular and Biological Investigations of Tritrichomonas foetus from Cattle, Domestic Cats and Pigs . (Thesis). University of Sydney. Retrieved from http://hdl.handle.net/2123/15168

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):

Morin-Adeline, Victoria. “Molecular and Biological Investigations of Tritrichomonas foetus from Cattle, Domestic Cats and Pigs .” 2016. Thesis, University of Sydney. Accessed April 11, 2021. http://hdl.handle.net/2123/15168.

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

MLA Handbook (7^th Edition):

Morin-Adeline, Victoria. “Molecular and Biological Investigations of Tritrichomonas foetus from Cattle, Domestic Cats and Pigs .” 2016. Web. 11 Apr 2021.

Vancouver:

Morin-Adeline V. Molecular and Biological Investigations of Tritrichomonas foetus from Cattle, Domestic Cats and Pigs . [Internet] [Thesis]. University of Sydney; 2016. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/2123/15168.

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

Council of Science Editors:

Morin-Adeline V. Molecular and Biological Investigations of Tritrichomonas foetus from Cattle, Domestic Cats and Pigs . [Thesis]. University of Sydney; 2016. Available from: http://hdl.handle.net/2123/15168

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

University of Texas – Austin

16. Rodríguez Mendoz, Álvaro Eugenio. Identifying mutations that enhance the evolutionary stability of fluorescent protein expression from a plasmid in Escherichia coli.

Degree: MA, Microbiology, 2014, University of Texas – Austin

URL: http://hdl.handle.net/2152/31999

► Synthetic biologists and metabolic engineers seek to design and create organisms with novel functions. A major difficulty with many designed genetic devices is that they… (more)

Subjects/Keywords: Evolutionary; Stability; Fluorescent; Protein; Mutations; Enhance

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Rodríguez Mendoz, �. E. (2014). Identifying mutations that enhance the evolutionary stability of fluorescent protein expression from a plasmid in Escherichia coli. (Masters Thesis). University of Texas – Austin. Retrieved from http://hdl.handle.net/2152/31999

Chicago Manual of Style (16^th Edition):

Rodríguez Mendoz, Álvaro Eugenio. “Identifying mutations that enhance the evolutionary stability of fluorescent protein expression from a plasmid in Escherichia coli.” 2014. Masters Thesis, University of Texas – Austin. Accessed April 11, 2021. http://hdl.handle.net/2152/31999.

MLA Handbook (7^th Edition):

Rodríguez Mendoz, Álvaro Eugenio. “Identifying mutations that enhance the evolutionary stability of fluorescent protein expression from a plasmid in Escherichia coli.” 2014. Web. 11 Apr 2021.

Vancouver:

Rodríguez Mendoz �E. Identifying mutations that enhance the evolutionary stability of fluorescent protein expression from a plasmid in Escherichia coli. [Internet] [Masters thesis]. University of Texas – Austin; 2014. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/2152/31999.

Council of Science Editors:

Rodríguez Mendoz �E. Identifying mutations that enhance the evolutionary stability of fluorescent protein expression from a plasmid in Escherichia coli. [Masters Thesis]. University of Texas – Austin; 2014. Available from: http://hdl.handle.net/2152/31999

Rice University

17. Pandey, Naresh. Characterizing the tolerance of near infrared fluorescent bacterial phytochromes to random backbone fission and circular permutation.

Degree: PhD, Natural Sciences, 2016, Rice University

URL: http://hdl.handle.net/1911/96201

► Protein fission, fusion, and circular permutation have been used to convert green fluorescent protein (GFP) family members into biosensors that dynamically report on cellular processes,… (more)

▼ Protein fission, fusion, and circular permutation have been used to convert green fluorescent protein (GFP) family members into biosensors that dynamically report on cellular processes, ranging from protein expression and metabolite concentrations to protein solubility, protein-protein interactions, and ligand-binding. Unfortunately, GFP are unsuitable for deep tissue reporting in animal models because the wavelengths of light used with these reporters is highly absorbed by tissues. In contrast, near infrared fluorescent protein (IFP and iRFP) reporters derived from bacterial phytochrome proteins (BphP) are excited by light in the near-infrared spectrum (~700 nm, less absorptive) and are better suited for probing cellular processes within tissues. IFP and iRFP can report on biological processes under anaerobic conditions because it uses biliverdin (BV) as a chromophore and does not require oxygen for maturation, a requisite for GFP maturation. Unlike GFP, IFP and iRFP are not yet able to report on wide-range of biological processes beyond gene expression. To report on gene expression, BphP must interlace its Per/ARNT/Sim (PAS) and cGMP phosphodiesterase/adenylcyclase/FhlA (GAF) domains into a topological knot. The extent to which this complex topology tolerates mutations (fission, fusion, and circular permutation) used to convert proteins into biosensors is not known. To better understand the tolerance of BphP to these types of mutational lesions, I have subjected IFP to random backbone fragmentation and iRFP to circular permutation using transposase mutagenesis. Screening a library of split IFP for fluorescent variants yielded thirteen unique fragmented IFP and with parent like spectral properties. These two-fragment IFP all required assistance from associating proteins for maximal fluorescence. These split sites displayed AND gate logic behavior when the ORFs encoding the different fragment are placed under distinct transcriptional regulation. In addition, screening a library of circularly permuted iRFP led to the discovery of twenty seven permuted iRFP variants with near infrared fluorescence. These variants arose from backbone fission in both the PAS and GAF domains, although the brightest permuted iRFP initiated at residues near the domain linker and termini. Biochemical analysis revealed that permuted iRFP display similar oligomerizatoin, quantum yield, and stability as native iRFP. These proteins also retained sufficient BV affinity serve as reporters of gene expression in mammalian cells without the addition of exogenous BV. The results described in this thesis represent the first study to map the tolerance of a BphP to random fragmentation and circular permutation. These results demonstrate that knotted BphP retain the ability to fold as their contact order changes, suggesting that these proteins can be further developed as reporters of biological processes like GFP. The split IFP represent a suite of assays that will be useful for monitoring the dynamics of a protein-protein interactions under conditions… Advisors/Committee Members: Silberg, Jonathan J (advisor).

Subjects/Keywords: near infrared fluorescent protein; circular permutation; knotted protein; protein engineering

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Pandey, N. (2016). Characterizing the tolerance of near infrared fluorescent bacterial phytochromes to random backbone fission and circular permutation. (Doctoral Dissertation). Rice University. Retrieved from http://hdl.handle.net/1911/96201

Chicago Manual of Style (16^th Edition):

Pandey, Naresh. “Characterizing the tolerance of near infrared fluorescent bacterial phytochromes to random backbone fission and circular permutation.” 2016. Doctoral Dissertation, Rice University. Accessed April 11, 2021. http://hdl.handle.net/1911/96201.

MLA Handbook (7^th Edition):

Pandey, Naresh. “Characterizing the tolerance of near infrared fluorescent bacterial phytochromes to random backbone fission and circular permutation.” 2016. Web. 11 Apr 2021.

Vancouver:

Pandey N. Characterizing the tolerance of near infrared fluorescent bacterial phytochromes to random backbone fission and circular permutation. [Internet] [Doctoral dissertation]. Rice University; 2016. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/1911/96201.

Council of Science Editors:

Pandey N. Characterizing the tolerance of near infrared fluorescent bacterial phytochromes to random backbone fission and circular permutation. [Doctoral Dissertation]. Rice University; 2016. Available from: http://hdl.handle.net/1911/96201

University of Alberta

18. Hoi, Hiofan. Development of monomeric fluorescent proteins and fluorescent protein-based biosensors.

Degree: PhD, Department of Chemistry, 2013, University of Alberta

URL: https://era.library.ualberta.ca/files/5712m723w

► Fluorescent protein (FP) technology is now an indispensible tool of biomedical research. Nevertheless, only a few members of the hundreds of existing FPs are generally… (more)

▼ Fluorescent protein (FP) technology is now an indispensible tool of biomedical research. Nevertheless, only a few members of the hundreds of existing FPs are generally regarded as the preferred options for most imaging applications. Accordingly, new FPs with ever-improved properties are in demand and worth pursuing, as these efforts may lead to variants that are closer to the “ideal” FP. Also there remain a tremendous number of opportunities for developing FP-based biosensors for probing biological process in vivo. This thesis describes our effort on engineering new FPs with improved properties, and further modifying them to create novel biosensors. Directed evolution and semi-rational protein engineering are the main techniques used to develop these new FPs and FP-based biosensors. The first class of FPs addressed in this thesis are the green-to-red photoconvertible FPs (pcFPs). In an effort to overcome the limitations imposed by the oligomeric structure of natural pcFPs, we created a new monomeric pcFP based on consensus design. Subsequent optimization yielded mClavGR2 and mMaple, two monomeric pcFPs displaying superior performance in folding and maturation, brightness, photoconversion efficiency and photostability. We demonstrate the application of mClavGR2 for dynamic monitoring of protein trafficking. Furthermore, in collaboration with researchers from several other groups, mMaple was demonstrated to be a multi-model probe that is suitable for use in several conventional and super-resolution fluorescence imaging modalities. Using mMaple as a template for single-FP biosensor design, we successfully combined the two most important implementations of FPs, the “highlightable” trait and the Ca2+ sensing capability, into one construct. Optimization, characterization and live cell imaging of the resulting green-to-red highlightable Ca2+ indicators are described. Another class of FPs that are of interest in this thesis are the true yellow emitting FPs that fill the spectral gap between monomeric greenish-yellow FPs and monomeric orange FPs. By disrupting the inter-subunit interfaces of zFP538, a FP with a distinct three-ring chromophore and an emission maximum at 538 nm, we successfully obtained its monomeric version and named it as mPapaya1. Again, characterization and live cell imaging application of mPapaya1 are described.

Subjects/Keywords: directed evolution; protein engineering; calcium indicator; genetically-encoded biosensor; fluorescent protein

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Hoi, H. (2013). Development of monomeric fluorescent proteins and fluorescent protein-based biosensors. (Doctoral Dissertation). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/5712m723w

Chicago Manual of Style (16^th Edition):

Hoi, Hiofan. “Development of monomeric fluorescent proteins and fluorescent protein-based biosensors.” 2013. Doctoral Dissertation, University of Alberta. Accessed April 11, 2021. https://era.library.ualberta.ca/files/5712m723w.

MLA Handbook (7^th Edition):

Hoi, Hiofan. “Development of monomeric fluorescent proteins and fluorescent protein-based biosensors.” 2013. Web. 11 Apr 2021.

Vancouver:

Hoi H. Development of monomeric fluorescent proteins and fluorescent protein-based biosensors. [Internet] [Doctoral dissertation]. University of Alberta; 2013. [cited 2021 Apr 11]. Available from: https://era.library.ualberta.ca/files/5712m723w.

Council of Science Editors:

Hoi H. Development of monomeric fluorescent proteins and fluorescent protein-based biosensors. [Doctoral Dissertation]. University of Alberta; 2013. Available from: https://era.library.ualberta.ca/files/5712m723w

19. Mythili, J Krishna. Identification of a fluorescent protein from a marine Zoanthid: Zoanthus Sansibaricus (Carlgren) from the intertidal rocky shore of Anjuna (Goa); -.

Degree: Pharmacy, 2011, Jawaharlal Nehru Technological University

URL: http://shodhganga.inflibnet.ac.in/handle/10603/4505

►

Zoanthids comprise an order of benthic, generally colonial Cnidarians, which can usually be distinguished from other hexacorallians by embedded sand and detritus in their mesoglea.… (more)

Subjects/Keywords: Green Fluorescent Protein; Chemistry; Protein; Zoanthid Taxonomy; Pharmacy

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Mythili, J. K. (2011). Identification of a fluorescent protein from a marine Zoanthid: Zoanthus Sansibaricus (Carlgren) from the intertidal rocky shore of Anjuna (Goa); -. (Thesis). Jawaharlal Nehru Technological University. Retrieved from http://shodhganga.inflibnet.ac.in/handle/10603/4505

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):

Mythili, J Krishna. “Identification of a fluorescent protein from a marine Zoanthid: Zoanthus Sansibaricus (Carlgren) from the intertidal rocky shore of Anjuna (Goa); -.” 2011. Thesis, Jawaharlal Nehru Technological University. Accessed April 11, 2021. http://shodhganga.inflibnet.ac.in/handle/10603/4505.

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

MLA Handbook (7^th Edition):

Mythili, J Krishna. “Identification of a fluorescent protein from a marine Zoanthid: Zoanthus Sansibaricus (Carlgren) from the intertidal rocky shore of Anjuna (Goa); -.” 2011. Web. 11 Apr 2021.

Vancouver:

Mythili JK. Identification of a fluorescent protein from a marine Zoanthid: Zoanthus Sansibaricus (Carlgren) from the intertidal rocky shore of Anjuna (Goa); -. [Internet] [Thesis]. Jawaharlal Nehru Technological University; 2011. [cited 2021 Apr 11]. Available from: http://shodhganga.inflibnet.ac.in/handle/10603/4505.

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

Council of Science Editors:

Mythili JK. Identification of a fluorescent protein from a marine Zoanthid: Zoanthus Sansibaricus (Carlgren) from the intertidal rocky shore of Anjuna (Goa); -. [Thesis]. Jawaharlal Nehru Technological University; 2011. Available from: http://shodhganga.inflibnet.ac.in/handle/10603/4505

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

University of California – San Francisco

20. Merksamer, Philip Ian. The Development and Application of Fluorescent Protein Reporters to Measure Endoplasmic Reticulum Stress in Single Cells.

Degree: Cell Biology, 2010, University of California – San Francisco

URL: http://www.escholarship.org/uc/item/1dw4d3hd

► In eukaryotic cells, secreted and membrane proteins fold within the endoplasmic reticulum (ER). Various physiological or pathophysiological conditions can disrupt ER protein folding homeostasis and… (more)

Subjects/Keywords: Cellular Biology; endoplasmic reticulum stress; fluorescent protein reporters; unfolded protein response

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Merksamer, P. I. (2010). The Development and Application of Fluorescent Protein Reporters to Measure Endoplasmic Reticulum Stress in Single Cells. (Thesis). University of California – San Francisco. Retrieved from http://www.escholarship.org/uc/item/1dw4d3hd

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):

Merksamer, Philip Ian. “The Development and Application of Fluorescent Protein Reporters to Measure Endoplasmic Reticulum Stress in Single Cells.” 2010. Thesis, University of California – San Francisco. Accessed April 11, 2021. http://www.escholarship.org/uc/item/1dw4d3hd.

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

MLA Handbook (7^th Edition):

Merksamer, Philip Ian. “The Development and Application of Fluorescent Protein Reporters to Measure Endoplasmic Reticulum Stress in Single Cells.” 2010. Web. 11 Apr 2021.

Vancouver:

Merksamer PI. The Development and Application of Fluorescent Protein Reporters to Measure Endoplasmic Reticulum Stress in Single Cells. [Internet] [Thesis]. University of California – San Francisco; 2010. [cited 2021 Apr 11]. Available from: http://www.escholarship.org/uc/item/1dw4d3hd.

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

Council of Science Editors:

Merksamer PI. The Development and Application of Fluorescent Protein Reporters to Measure Endoplasmic Reticulum Stress in Single Cells. [Thesis]. University of California – San Francisco; 2010. Available from: http://www.escholarship.org/uc/item/1dw4d3hd

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

University of New Mexico

21. Phillips, Genevieve Kate. DEVELOPING FLUOROGEN ACTIVATING PROTEIN-FLUORESCENT PROTEIN FRET PAIRS FOR LIVE CELL IMAGING.

Degree: Biomedical Sciences Graduate Program, 2017, University of New Mexico

URL: https://digitalrepository.unm.edu/biom_etds/161

► Fluorogen activating proteins (FAPs) are genetically encoded tags made from single chain antibody fragments (scFv) designed to bind fluorogens with high specificity. Both the… (more)

▼ Fluorogen activating proteins (FAPs) are genetically encoded tags made from single chain antibody fragments (scFv) designed to bind fluorogens with high specificity. Both the fluorogen and FAP can be modified to provide flexibility in properties such as affinity, membrane permeability, spectra, and quantum yield. The fluorogen Malachite Green (MG) has two excitation peaks, the maximum at 630 nm and a secondary peak at 450 nm. The emission spectra of blue-emitting fluorescence proteins, such as mCerulean (mCer), overlap with the MG secondary peak, generating a FRET pair with large Stokes shift emission. Using 405 nm excitation of mCer, we observe acceptor sensitized emission at wavelengths greater than 650 nm with no spectral crosstalk between the donor and acceptor channels. Additionally, donor only controls can be acquired for all cells as the acceptor is not present until after the addition of the fluorogen, providing intra-cellular control. The FAP-FRET system has been characterized using proof of principle constructs: FAP-mCer-transmembrane (TM) as a positive FRET control and FAP-TM-mCer as a negative FRET control and expressed in HeLa cells. Multiple MG derivatives were compared and imaging parameters were optimized to determine the optimal FRET pair. Analysis was performed using code written in Matlab to mask the cell membrane and quantify FRET efficiencies, based on donor intensity before and after addition of fluorogen. Data from several fluorogen showed high energy transfer efficiency (~30%) with the FAP-mCer-TM construct compared to negligible FRET (~4%) for FAP-TM-mCer. Additional techniques were performed to support the FRET efficiency data, including spectral imaging and FLIM, which also reported FRET efficiency around 30% with the positive constructs and negligible FRET with the negative constructs. The FAP-FRET system is currently being used to study the kinetics of signaling proteins within the FcεRI pathway. Advisors/Committee Members: Dr. Diane Lidke, Dr. Angela Wandinger-Ness, Dr. Heather Ward, Dr. Bill Shuttleworth.

Subjects/Keywords: Fluorogen activating protein (FAP); FRET; Fluorescent protein; Medicine and Health Sciences

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Phillips, G. K. (2017). DEVELOPING FLUOROGEN ACTIVATING PROTEIN-FLUORESCENT PROTEIN FRET PAIRS FOR LIVE CELL IMAGING. (Masters Thesis). University of New Mexico. Retrieved from https://digitalrepository.unm.edu/biom_etds/161

Chicago Manual of Style (16^th Edition):

Phillips, Genevieve Kate. “DEVELOPING FLUOROGEN ACTIVATING PROTEIN-FLUORESCENT PROTEIN FRET PAIRS FOR LIVE CELL IMAGING.” 2017. Masters Thesis, University of New Mexico. Accessed April 11, 2021. https://digitalrepository.unm.edu/biom_etds/161.

MLA Handbook (7^th Edition):

Phillips, Genevieve Kate. “DEVELOPING FLUOROGEN ACTIVATING PROTEIN-FLUORESCENT PROTEIN FRET PAIRS FOR LIVE CELL IMAGING.” 2017. Web. 11 Apr 2021.

Vancouver:

Phillips GK. DEVELOPING FLUOROGEN ACTIVATING PROTEIN-FLUORESCENT PROTEIN FRET PAIRS FOR LIVE CELL IMAGING. [Internet] [Masters thesis]. University of New Mexico; 2017. [cited 2021 Apr 11]. Available from: https://digitalrepository.unm.edu/biom_etds/161.

Council of Science Editors:

Phillips GK. DEVELOPING FLUOROGEN ACTIVATING PROTEIN-FLUORESCENT PROTEIN FRET PAIRS FOR LIVE CELL IMAGING. [Masters Thesis]. University of New Mexico; 2017. Available from: https://digitalrepository.unm.edu/biom_etds/161

University of Edinburgh

22. Chen, Kai. Use of green fluorescent protein for the analysis of protein-protein and protein-DNA interactions.

Degree: PhD, 2011, University of Edinburgh

URL: http://hdl.handle.net/1842/4886

► Restriction modification (RM) systems play a crucial role in preventing the entry of foreign DNA into the bacterial cell. The best studied Type I RM… (more)

▼ Restriction modification (RM) systems play a crucial role in preventing the entry of foreign DNA into the bacterial cell. The best studied Type I RM system is EcoKI from Escherichia coli K12. Both bacteriophage and conjugative plasmids have developed a variety of strategies to circumvent the host RM system. One such strategy involves the production of antirestriction proteins that mimic a short segment of DNA and efficiently inhibit the RM system. The main aim of this project was to analyse the interaction of EcoKI and its cognate methylase (MTase) with the T7 antirestriction protein, known as overcome classical restriction (Ocr), and various ArdA antirestriction proteins. Currently, there is a paucity of structural data on the complex formed between the Type I system and the antirestriction proteins. The aim of this work was twofold; (i) compare the interaction of MTase with DNA and Ocr and (ii) quantify the strength of interaction between MTase and various ArdA proteins. The MTase was fused to the Green Fluorescent Protein (GFP) to facilitate determination of the orientation of interaction with DNA and Ocr. Time resolved fluorescence measurements were carried out using the GFP-MTase fusion to determine the fluorescence lifetime and anisotropy decay. These experiments were conducted using a time resolved fluorescence instrument fabricated in-house. The values determined in these experiments were then used to perform fluorescence resonance energy transfer (FRET) measurements with fluorescently labelled DNA or Ocr. These measurements gave information concerning the relative orientation of the MTase with either DNA or Ocr. The GFP-MTase fusion was also used to quantify the strength of interaction with various ArdA proteins. Previous attempts to determine the strength of interaction between MTase and ArdA proteins by employing conventional techniques have been unsuccessful. Therefore, a novel method was developed that exploits the interaction of MTase with a cation exchange medium, which can subsequently be displaced upon binding to ArdA. This method facilitated the determination, for the first time, of a set of binding affinities for the MTase and ArdA interaction.

Subjects/Keywords: 572.8; GFP; green fluorescent protein; protein-protein; DNA restriction/modification; Time-resolved fluorescence

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Chen, K. (2011). Use of green fluorescent protein for the analysis of protein-protein and protein-DNA interactions. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/4886

Chicago Manual of Style (16^th Edition):

Chen, Kai. “Use of green fluorescent protein for the analysis of protein-protein and protein-DNA interactions.” 2011. Doctoral Dissertation, University of Edinburgh. Accessed April 11, 2021. http://hdl.handle.net/1842/4886.

MLA Handbook (7^th Edition):

Chen, Kai. “Use of green fluorescent protein for the analysis of protein-protein and protein-DNA interactions.” 2011. Web. 11 Apr 2021.

Vancouver:

Chen K. Use of green fluorescent protein for the analysis of protein-protein and protein-DNA interactions. [Internet] [Doctoral dissertation]. University of Edinburgh; 2011. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/1842/4886.

Council of Science Editors:

Chen K. Use of green fluorescent protein for the analysis of protein-protein and protein-DNA interactions. [Doctoral Dissertation]. University of Edinburgh; 2011. Available from: http://hdl.handle.net/1842/4886

Georgia Tech

23. Fellows, William Brett. Combinatorial synthesis of new GFP- and RFP-like chromophores and their photophysical properties.

Degree: PhD, Chemistry and Biochemistry, 2014, Georgia Tech

URL: http://hdl.handle.net/1853/52318

► A new synthetic methodology for the combinatorial preparation of C-terminus-modified Green and Red Fluorescent Protein chromophores is described. This method involves the modification of the… (more)

Subjects/Keywords: Green fluorescent protein; Chromophore; Synthesis; Combinatorial; Red fluorescent protein; Hot dog stacking; Solid state; Crystal structure; Aggregate induced emission; Reprecipitation

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Fellows, W. B. (2014). Combinatorial synthesis of new GFP- and RFP-like chromophores and their photophysical properties. (Doctoral Dissertation). Georgia Tech. Retrieved from http://hdl.handle.net/1853/52318

Chicago Manual of Style (16^th Edition):

Fellows, William Brett. “Combinatorial synthesis of new GFP- and RFP-like chromophores and their photophysical properties.” 2014. Doctoral Dissertation, Georgia Tech. Accessed April 11, 2021. http://hdl.handle.net/1853/52318.

MLA Handbook (7^th Edition):

Fellows, William Brett. “Combinatorial synthesis of new GFP- and RFP-like chromophores and their photophysical properties.” 2014. Web. 11 Apr 2021.

Vancouver:

Fellows WB. Combinatorial synthesis of new GFP- and RFP-like chromophores and their photophysical properties. [Internet] [Doctoral dissertation]. Georgia Tech; 2014. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/1853/52318.

Council of Science Editors:

Fellows WB. Combinatorial synthesis of new GFP- and RFP-like chromophores and their photophysical properties. [Doctoral Dissertation]. Georgia Tech; 2014. Available from: http://hdl.handle.net/1853/52318

University of Tennessee – Knoxville

24. Rice, John Hollis. Bioconfinement of a putatively sterile Nicotiana hybrid and development of tools for assessing gene flow.

Degree: MS, Plant Sciences, 2013, University of Tennessee – Knoxville

URL: https://trace.tennessee.edu/utk_gradthes/2487

► Production of transgenic crops in open field environments is an ongoing concern of due to the potential for gene flow. New transgenic crops, such… (more)

▼ Production of transgenic crops in open field environments is an ongoing concern of due to the potential for gene flow. New transgenic crops, such as plant-made-pharmaceuticals may generate additional concerns about effects of adventitious transgenes. Use of a bioconfinement strategy may alleviate any consequences by preventing gene flow. The following chapters discuss previous and current research on gene flow, testing of a Nicotiana hybrid system for bioconfinement efficiency, and development of methods for transgene detection. The candidate ‘platform plant’ that was tested is a Nicotiana hybrid (Nicotiana tabacum ‘TN 90’ × Nicotiana glauca) previously identified to be sexually sterile. To quantify gene flow from hybrids, the mGFP5ER gene encoding for green fluorescent protein (GFP) was inserted into the paternal lines, which were crossed to form the hybrid. The DNA content and male fertility of these lines were used to characterize GFP-tagged hybrid lines. There were no differences in DNA content but significant differences in male fertility, in which pollen germination was observed at low rates. Two field gene flow studies revealed GFP-hybrids were not totally sterile since hybrids outcrossed and were pollinated by N. tabacum pollen, but they produced few viable seed. These results were confirmed with manual greenhouse crosses. Biomass studies revealed that the GFP-hybrids were comparable in productivity to an N. tabacum cultivar typically used in field production. An additional tagging strategy was created to produce the orange fluorescent protein (OFP), tdTomato-ER, in pollen by using pollen specific promoters in addition to the whole plant GFP cassettes. N. tabacum ‘TN 90’ and N. glauca were transformed, bred and hybridized to generate a hybrid that produced pollen tagged with orange fluorescent protein. Manual crosses were performed in a greenhouse and partially similar results were obtained compared with previous GFP-tagged hybrid crosses. OFP-tagged hybrid outcrossing produced totally non-viable seed and when non-transgenic N. tabacum was supplied as a pollen donor to the OFP-tagged hybrids some non-tagged progeny were observed. The results suggest the Nicotiana hybrid could be a productive biomanufacturing platform and could provide total bioconfinement if grown in physical isolation from N. tabacum and N. glauca. Advisors/Committee Members: C. Neal Stewart Jr., Charles Kwit, Randall L. Small.

Subjects/Keywords: bioconfinement; Nicotiana; green fluorescent protein; orange fluorescent protein; gene flow; plant molecular farming; Biotechnology; Molecular Biology; Plant Breeding and Genetics

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Rice, J. H. (2013). Bioconfinement of a putatively sterile Nicotiana hybrid and development of tools for assessing gene flow. (Thesis). University of Tennessee – Knoxville. Retrieved from https://trace.tennessee.edu/utk_gradthes/2487

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):

Rice, John Hollis. “Bioconfinement of a putatively sterile Nicotiana hybrid and development of tools for assessing gene flow.” 2013. Thesis, University of Tennessee – Knoxville. Accessed April 11, 2021. https://trace.tennessee.edu/utk_gradthes/2487.

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

MLA Handbook (7^th Edition):

Rice, John Hollis. “Bioconfinement of a putatively sterile Nicotiana hybrid and development of tools for assessing gene flow.” 2013. Web. 11 Apr 2021.

Vancouver:

Rice JH. Bioconfinement of a putatively sterile Nicotiana hybrid and development of tools for assessing gene flow. [Internet] [Thesis]. University of Tennessee – Knoxville; 2013. [cited 2021 Apr 11]. Available from: https://trace.tennessee.edu/utk_gradthes/2487.

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

Council of Science Editors:

Rice JH. Bioconfinement of a putatively sterile Nicotiana hybrid and development of tools for assessing gene flow. [Thesis]. University of Tennessee – Knoxville; 2013. Available from: https://trace.tennessee.edu/utk_gradthes/2487

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

Humboldt State University

25. Cramer, Philip B. Using transgenic Caenorhabditis elegans as a bioindicator: a useful tool for examining potential environmental hazards, or just a bunch of glowing worms?.

Degree: MS, Biology, 2012, Humboldt State University

URL: http://hdl.handle.net/2148/1260

► A total of 17 transgenic strains of Caenorhabditis elegans were evaluated for use as bioindicators for the presence of toxins in an environmental sample. Every… (more)

▼ A total of 17 transgenic strains of Caenorhabditis elegans were evaluated for use as bioindicators for the presence of toxins in an environmental sample. Every strain used had Green Fluorescent Protein (GFP) production driven by the promoter region of a gene known to be involved in stress response, metabolism, or development. Controlled exposures were performed on each strain using 22 environmental toxins and contaminants. Changes in GFP intensity between exposed groups and controls were considered proof of changes in that genes expression pattern as a direct result of exposure to the toxin. The goal was to construct expression profiles for each strain using the chosen toxins. The strains could then be used to evaluate environmental samples for the presence of unknown toxins with the expression profiles serving as a preliminary indicator of the amount and type of toxin present. The metallothionein-linked strain CL 2122 showed significant upregulation in the Mtl-2 gene following exposure to 10, 50, and 100 ??M cadmium, copper, and lead. This strain also exhibited significant downregulation of metallothionein expression following exposure to diazinon, hexazinone, iron (II) sulfate, paraquat, and piperonyl butoxide, which exemplifies how even a metals-specific stress response can be altered by certain toxins to yield difficult to interpret results. Initially it was thought that a panel of 6-8 strains could be formed that showed clear expression profiles to common environmental pollutants and used to examine samples that had unknown toxins. It was envisioned that the panel would have at least 2 strains that are metals-responsive, two strains that are general organic toxin-responsive, and two strains that are endocrine disruptor and/or estrogen sensitive. This proved to be unrealistic for the genes examined as no strain yielded results that were totally unambiguous. The results indicated that single-gene biomarker-based bioassays could only be used in limited situations, such as large single-toxin releases, or when identifying the exact toxin is not as important as assessing the overall safety of the sample in question. Advisors/Committee Members: O'Gara, Bruce A..

Subjects/Keywords: Caenorhabditis; Elegans; Biomarker; Bioindicator; Transgenic; Green fluorescent protein; GFP

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Cramer, P. B. (2012). Using transgenic Caenorhabditis elegans as a bioindicator: a useful tool for examining potential environmental hazards, or just a bunch of glowing worms?. (Masters Thesis). Humboldt State University. Retrieved from http://hdl.handle.net/2148/1260

Chicago Manual of Style (16^th Edition):

Cramer, Philip B. “Using transgenic Caenorhabditis elegans as a bioindicator: a useful tool for examining potential environmental hazards, or just a bunch of glowing worms?.” 2012. Masters Thesis, Humboldt State University. Accessed April 11, 2021. http://hdl.handle.net/2148/1260.

MLA Handbook (7^th Edition):

Cramer, Philip B. “Using transgenic Caenorhabditis elegans as a bioindicator: a useful tool for examining potential environmental hazards, or just a bunch of glowing worms?.” 2012. Web. 11 Apr 2021.

Vancouver:

Cramer PB. Using transgenic Caenorhabditis elegans as a bioindicator: a useful tool for examining potential environmental hazards, or just a bunch of glowing worms?. [Internet] [Masters thesis]. Humboldt State University; 2012. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/2148/1260.

Council of Science Editors:

Cramer PB. Using transgenic Caenorhabditis elegans as a bioindicator: a useful tool for examining potential environmental hazards, or just a bunch of glowing worms?. [Masters Thesis]. Humboldt State University; 2012. Available from: http://hdl.handle.net/2148/1260

University of California – Riverside

26. Fan, Yichong. Expanding the Current Imaging Toolkit With Novel Fluorescent Protein Based Biosensors.

Degree: Environmental Toxicology, 2017, University of California – Riverside

URL: http://www.escholarship.org/uc/item/627146dj

► The objective of my Ph.D. study is to develop novel genetically encoded fluorescent biosensors to image and dissect biological signaling pathways in the context of… (more)

▼ The objective of my Ph.D. study is to develop novel genetically encoded fluorescent biosensors to image and dissect biological signaling pathways in the context of live cells. I utilized protein engineering techniques to convert fluorescent proteins into fluorescent biosensors that can actively respond to specific, spatiotemporally organized cellular changes.In this thesis, we expanded the fluorescent protein toolkit by engineering one of the first red fluorescent probes⎯rxRFP1⎯for sensing general redox states in the live cells. To further extend the usage of this sensor in various subcellular domains, such as mitochondria, endoplasmic reticulum, and the cell nucleus, we developed a group of rxRFP1 mutants showing different midpoint redox potentials for studying compartmentalized redox dynamics under various pathophysiological conditions. We also developed the first genetically encoded fluorescent biosensor for thioredoxin (Trx) redox by engineering a redox relay between the active-site cysteines of human Trx1 and rxRFP1. We utilized the resultant biosensor⎯TrxRFP1⎯to selectively monitor the perturbations of Trx redox in various mammalian cell lines. We further combined TrxRFP1 with a green fluorescent Grx1-roGFP2 biosensor to simultaneously monitor the dynamics of the two major cellular antioxidant systems, Trx and glutathione, in live cells in response to chemically and physiologically relevant stimuli. We exploit another strategy which introduces reactive functional groups into circular permutated fluorescent proteins (cpFPs) using a genetic code expansion technology. Through a powerful directed protein evolution process, we were able to modulate the reactivity and chemoselectivity of an introduced p-boronophenylalanine (pBoF) in a cpRFP scaffold, resulting in fluorescent probes selectively responsive to hydrogen peroxide (H2O2) and peroxynitrite (ONOO—). Furthermore, by using boronic acid and short peptides as synergistic recognition motifs, we were able to engineer a series of reversible probes for nucleotides and carbohydrates showing surprisingly high specificity and large dynamic ranges. We have successfully utilized this new family of fluorescent probes to visualize various cellular activities.

Subjects/Keywords: Toxicology; biological processes; biosensor; fluorescent protein; imaging; redox signaling

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Fan, Y. (2017). Expanding the Current Imaging Toolkit With Novel Fluorescent Protein Based Biosensors. (Thesis). University of California – Riverside. Retrieved from http://www.escholarship.org/uc/item/627146dj

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):

Fan, Yichong. “Expanding the Current Imaging Toolkit With Novel Fluorescent Protein Based Biosensors.” 2017. Thesis, University of California – Riverside. Accessed April 11, 2021. http://www.escholarship.org/uc/item/627146dj.

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

MLA Handbook (7^th Edition):

Fan, Yichong. “Expanding the Current Imaging Toolkit With Novel Fluorescent Protein Based Biosensors.” 2017. Web. 11 Apr 2021.

Vancouver:

Fan Y. Expanding the Current Imaging Toolkit With Novel Fluorescent Protein Based Biosensors. [Internet] [Thesis]. University of California – Riverside; 2017. [cited 2021 Apr 11]. Available from: http://www.escholarship.org/uc/item/627146dj.

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

Council of Science Editors:

Fan Y. Expanding the Current Imaging Toolkit With Novel Fluorescent Protein Based Biosensors. [Thesis]. University of California – Riverside; 2017. Available from: http://www.escholarship.org/uc/item/627146dj

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

University of Alberta

27. Wu, Jiahui. Development of red fluorescent protein-based calcium ion and glutamate indicators.

Degree: PhD, Department of Chemistry, 2014, University of Alberta

URL: https://era.library.ualberta.ca/files/c9s1616409

► The discovery and subsequent applications of fluorescent proteins (FPs) launched a new era for live cell fluorescence imaging. The design and developments of FP-based indicators… (more)

▼ The discovery and subsequent applications of fluorescent proteins (FPs) launched a new era for live cell fluorescence imaging. The design and developments of FP-based indicators have further solidified the versatility of FPs and rendered them as indispensable tools in life science research now more than ever. Despite the tremendous developments and efforts invested in the field of FP-based indicators, there remain numerous opportunities in engineering indicators with improved or novel properties for studying biological processes in vivo. In this thesis we describe our efforts in developing a series of FP-based calcium ion (Ca2+) and glutamate indicators with various colors and useful spectral properties as versatile tools for interrogating cell signaling in cell biology. In this thesis, we first describe our efforts in employing protein engineering to expand the color palette of genetically encoded Ca2+ indicators to include intensiometric orange, improved red and ratiometric red fluorescent variants. We demonstrate these new indicators' utility by performing Ca2+ imaging in cultured human cell lines, slice culture of developing mouse neocortex, organotypic hippocampal slice cultures and the visual system of albino tadpoles. Using our intensiometric red Ca2+ indicators, R-GECO1 and R-GECO1.2, as templates, we further engineered a series of low affinity R-GECOs with dissociation constants (Kds) ranging from 12 µM to more than 540 µM. We demonstrate that these indicators can be used to image cell compartments with high Ca2+ concentration or with a broad range of Ca2+ change, such as the endoplasmic reticulum (ER) and mitochondria. We also demonstrate these new red Ca2+ indicators with low affinities can be used to monitor ER and mitochondrial Ca2+ in combination with a green fluorescent protein (GFP)-based reporter. We also report in this thesis the development, optimization and characterization of the first red fluorescent protein (RFP)-based glutamate indicator, GltR1. We demonstrate GltR1 can detect glutamate changes on the surface of cultured human cells, as well as the glutamate dynamics during spontaneous activities of dissociated rat hippocampal neurons.

Subjects/Keywords: fluorescent protein; GECO; biosensor; glutamate indicator; fluorescence Ca2+ imaging

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Wu, J. (2014). Development of red fluorescent protein-based calcium ion and glutamate indicators. (Doctoral Dissertation). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/c9s1616409

Chicago Manual of Style (16^th Edition):

Wu, Jiahui. “Development of red fluorescent protein-based calcium ion and glutamate indicators.” 2014. Doctoral Dissertation, University of Alberta. Accessed April 11, 2021. https://era.library.ualberta.ca/files/c9s1616409.

MLA Handbook (7^th Edition):

Wu, Jiahui. “Development of red fluorescent protein-based calcium ion and glutamate indicators.” 2014. Web. 11 Apr 2021.

Vancouver:

Wu J. Development of red fluorescent protein-based calcium ion and glutamate indicators. [Internet] [Doctoral dissertation]. University of Alberta; 2014. [cited 2021 Apr 11]. Available from: https://era.library.ualberta.ca/files/c9s1616409.

Council of Science Editors:

Wu J. Development of red fluorescent protein-based calcium ion and glutamate indicators. [Doctoral Dissertation]. University of Alberta; 2014. Available from: https://era.library.ualberta.ca/files/c9s1616409

University of Georgia

28. Majsztrik, John Christopher. Subnuclear localization and interaction of selected Aux/IAA and ARF proteins in vivo.

Degree: 2014, University of Georgia

URL: http://hdl.handle.net/10724/21898

► The plant hormone auxin has been studied for many years and is instrumental for plant growth and development. Auxin regulates gene transcription of at least… (more)

Subjects/Keywords: Auxin Response Factor (ARF); Aux/IAA; Fluorescent protein; Nuclear localization

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Majsztrik, J. C. (2014). Subnuclear localization and interaction of selected Aux/IAA and ARF proteins in vivo. (Thesis). University of Georgia. Retrieved from http://hdl.handle.net/10724/21898

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):

Majsztrik, John Christopher. “Subnuclear localization and interaction of selected Aux/IAA and ARF proteins in vivo.” 2014. Thesis, University of Georgia. Accessed April 11, 2021. http://hdl.handle.net/10724/21898.

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

MLA Handbook (7^th Edition):

Majsztrik, John Christopher. “Subnuclear localization and interaction of selected Aux/IAA and ARF proteins in vivo.” 2014. Web. 11 Apr 2021.

Vancouver:

Majsztrik JC. Subnuclear localization and interaction of selected Aux/IAA and ARF proteins in vivo. [Internet] [Thesis]. University of Georgia; 2014. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/10724/21898.

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

Council of Science Editors:

Majsztrik JC. Subnuclear localization and interaction of selected Aux/IAA and ARF proteins in vivo. [Thesis]. University of Georgia; 2014. Available from: http://hdl.handle.net/10724/21898

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

McMaster University

29. Kuryllo, Kacper. Detecting RNA Regulatory Interactions in Bacterial Cells.

Degree: PhD, 2014, McMaster University

URL: http://hdl.handle.net/11375/16296

►

Non-coding RNAs are involved in the regulation of most major cellular process in Escherichia coli. With current technologies, many of these molecules have been identified;… (more)

Subjects/Keywords: RNA; gene regulation; sRNA; riboswitch; biosensor; fluorescent protein; GFP; Escherichia coli

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Kuryllo, K. (2014). Detecting RNA Regulatory Interactions in Bacterial Cells. (Doctoral Dissertation). McMaster University. Retrieved from http://hdl.handle.net/11375/16296

Chicago Manual of Style (16^th Edition):

Kuryllo, Kacper. “Detecting RNA Regulatory Interactions in Bacterial Cells.” 2014. Doctoral Dissertation, McMaster University. Accessed April 11, 2021. http://hdl.handle.net/11375/16296.

MLA Handbook (7^th Edition):

Kuryllo, Kacper. “Detecting RNA Regulatory Interactions in Bacterial Cells.” 2014. Web. 11 Apr 2021.

Vancouver:

Kuryllo K. Detecting RNA Regulatory Interactions in Bacterial Cells. [Internet] [Doctoral dissertation]. McMaster University; 2014. [cited 2021 Apr 11]. Available from: http://hdl.handle.net/11375/16296.

Council of Science Editors:

Kuryllo K. Detecting RNA Regulatory Interactions in Bacterial Cells. [Doctoral Dissertation]. McMaster University; 2014. Available from: http://hdl.handle.net/11375/16296

30. Drufva, Erin. DEVELOPMENT OF PROTEIN DISPLAY SYSTEMS AND GENETIC TOOLS FOR SPORE-FORMING BACTERIA.

Degree: PhD, 2018, University of New Hampshire

URL: https://scholars.unh.edu/dissertation/2398

► One major area of synthetic biology is to engineer microbial cells and subcellular systems for diverse applications including biosynthesis, biocatalysis, therapeutics, drug delivery, and… (more)

▼ One major area of synthetic biology is to engineer microbial cells and subcellular systems for diverse applications including biosynthesis, biocatalysis, therapeutics, drug delivery, and bioremediation. For most applications, robust cellular systems are preferred for longer activity half-life and resistance to harsh environments. Two projects related to robust cellular systems involving Gram-positive bacteria are presented in this work. One is to develop thermostable genetic reporters for Geobacilli species and the other is to display an enzyme on the Bacillus subtilis spore surface to enhance its robustness and present an alternative to purified enzymes for industrial applications. Bacillus subtilis and Geobacillus thermoglucosidans are gram-positive, spore-forming bacteria. They secrete many proteins used industrially for the production of paper, food, textiles, chemicals, medicine, and cosmetics. Since G. thermoglucosidans is thermostable with an optimal growth temperature of 60ºC, its secreted proteins are also thermostable which proves advantageous for a variety of industrial applications. Additionally, a strain of G. thermoglucosidans has been used for the production of ethanol from biomass. Unfortunately the inner workings of G. thermoglucosidans are still poorly understood and a genetic toolkit is necessary to better discover how to improve them via genetic engineering for industrial use. Important components of this toolkit are genetic reporters which allow for the analysis of gene expression in G. thermoglucosidans. Fluorescent proteins are commonly used reporters for other bacterial species due to their easily observed and readily measured signal, however no thermostable fluorescent proteins have been shown to be functional in Geobacillus. Seven different fluorescent proteins including mCherry, Venus, GFP, sfGFP, GFPmut3, mCherry (Gt), and Venus (Gt) were tested for stability and functionality in Geobacillus thermoglucosidans. Venus (Gt) and mCherry (Gt) were codon optimized for this bacterium with the goal of increasing expression level and thus improving the fluorescence signal. The fluorescence intensity of each fluorescent protein expressed in G. thermoglucosidans was measured after several hours of bacterial growth at 50ºC and 60ºC. Venus, mCherry, Venus (Gt), mCherry (Gt), and sfGFP all had signal when expressed in G. thermoglucosidans at 50ºC and sfGFP had signal at 60ºC. Therefore, fluorescent reporter proteins in three different colors were found to be functional in G. thermoglucosidans. This will further genetic engineering of the species for thermostable protein production, bioremediation, and biofuel production. Bacillus subtilis is Generally Regarded as Safe (GRAS) by the FDA and amenable toward genetic manipulation. Thus it has been engineered for the production of many heterologous proteins. Oftentimes, proteins secreted by bacteria are purified for industrial use. However, protein purification is expensive and time-consuming and long-term storage of purified proteins… Advisors/Committee Members: Kang Wu, Kang Wu, Russell Carr.

Subjects/Keywords: Bacillus; fluorescent protein; Geobacillus; immobilize; laccase; spore; Microbiology; Biology; Chemical engineering

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Drufva, E. (2018). DEVELOPMENT OF PROTEIN DISPLAY SYSTEMS AND GENETIC TOOLS FOR SPORE-FORMING BACTERIA. (Doctoral Dissertation). University of New Hampshire. Retrieved from https://scholars.unh.edu/dissertation/2398

Chicago Manual of Style (16^th Edition):

Drufva, Erin. “DEVELOPMENT OF PROTEIN DISPLAY SYSTEMS AND GENETIC TOOLS FOR SPORE-FORMING BACTERIA.” 2018. Doctoral Dissertation, University of New Hampshire. Accessed April 11, 2021. https://scholars.unh.edu/dissertation/2398.

MLA Handbook (7^th Edition):

Drufva, Erin. “DEVELOPMENT OF PROTEIN DISPLAY SYSTEMS AND GENETIC TOOLS FOR SPORE-FORMING BACTERIA.” 2018. Web. 11 Apr 2021.

Vancouver:

Drufva E. DEVELOPMENT OF PROTEIN DISPLAY SYSTEMS AND GENETIC TOOLS FOR SPORE-FORMING BACTERIA. [Internet] [Doctoral dissertation]. University of New Hampshire; 2018. [cited 2021 Apr 11]. Available from: https://scholars.unh.edu/dissertation/2398.

Council of Science Editors:

Drufva E. DEVELOPMENT OF PROTEIN DISPLAY SYSTEMS AND GENETIC TOOLS FOR SPORE-FORMING BACTERIA. [Doctoral Dissertation]. University of New Hampshire; 2018. Available from: https://scholars.unh.edu/dissertation/2398

		in
And / Or
		in
And / Or
		in
And / Or
		in

Open Access Theses and Dissertations