Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
Results
Showing 1 to 18 of 18 for “"intein"”.
-
A Selection for Intein Splicing inPhage Assisted Continuous Evolution
… goal in chemical biology. Trans-splicing inteins fulfill a crucial niche in this area by enabling post translational splicing of separate polypeptides, introduction of post translational modifications, and other chemistry on the amide backbone. This has motivated engineering and evolution …
-
Split Inteins: From Mechanistic Studies to Novel Protein Engineering Technologies
<p>Inteins are auto-processing protein domains that carry out a post-translational process known as protein splicing. This process is characterized by excision of the intein (intervening protein) domain from within a larger polypeptide sequence with concomitant ligation of the flanking extein ( …
-
Mobile genetic elements in Coxiella burnetii: Friends, foes or just indifferent?
… (IVS) that interrupt the 23S rRNA gene; an intein within dnaB (encoding replicative DNA helicase) and a homing endonuclease. The introns are self-splicing ribozymes and able to inhibit ribosome function and retard bacterial growth through internal guide sequence (IGS)-dependent and …
-
Mobile genetic elements in Coxiella burnetii: Friends, foes or just indifferent?
… (IVS) that interrupt the 23S rRNA gene; an intein within dnaB (encoding replicative DNA helicase) and a homing endonuclease. The introns are self-splicing ribozymes and able to inhibit ribosome function and retard bacterial growth through internal guide sequence (IGS)-dependent and …
-
Structure-Guided Development of Antifungal Protein Farnesyltransferase Inhibitors and DNA Polymerase Engineering
… inhibitor development. In the second study, an intein was inserted into DNA polymerases to produce temperature controlled enzymes. The intein controlled DNA polymerases only showed activities after intein splicing triggered by high temperature (>60oC), enabling the capability of conducting …
-
Protein Semi-Synthesis in Vivo
… is genetically tagged with one half of a split-intein. The complementary half is linked in vitro to the synthetic probe and this fusion is delivered into cells using a transduction peptide. Association of the intein halves in the cytosol triggers protein transsplicing, resulting in the ligation …
-
Structure and regulation of yeast glycogen synthase
… crystallography, site-directed mutagenesis, intein-mediated peptide ligation and enzyme assays. We have solved the tetrameric structure of the yeast enzyme in two different activity states; the resting enzyme and the activated state when complexed with glucose-6-phosphate. Binding of …
-
Conditional Protein Alleles and Regulation of Bacterial Transcription: A Thesis in Two Acts
… we term conditional protein splicing (CPS), an intein is split into two inactive fragments which are activated by heterodimerization. By fragmenting the target protein and fusing the pieces to this split intein, we were able to control the target protein's activity through splicing. In the …
-
Development of Bacillus subtilis spores and cells for surface display of proteins
… on the spore surface, separated by a mini-intein derived from the SSp DnaB, which was then used as self-cleaving tag for purification by shifting the pH and/or temperature conditions, with no addition of any proteases or thiol reagent (Mathys et al., 1999). To construct the system, the …
-
ADVANCED AAV-MEDIATED LIVER-DIRECTED GENE THERAPIES FOR HAEMOPHILIA A AND MUCOPOLYSACCHARIDOSIS TYPE VI
… trans-splicing (PTS) strategy utilizing split inteins. PTS is a natural mechanism employed by organisms across all domains of life to reconstitute large proteins from shorter precursors. Split inteins facilitate this process by selfexcising from the host protein in an energy-independent manner. …
-
Mechanistic investigations of the radical initiation pathway of class I ribonucleotide reductase from Escherichia coli
… an effort to study radical initiation, we used intein technology to semi-synthetically synthesize R2 (375 residues) where Y356 is replaced with an unnatural amino acid. Thus far, we have replaced Y356 with 3- nitrotyrosine (NO2Y), 2,3-difluorotyrosine (2,3-DFY), 2,3,5-trifluorotyrosine …
-
Allosteric Determinants of Guanine Nucleotide Binding Proteins and Methods to Crystallize the Cytosolic Domains of Adenylyl Cyclase
… but failed to diffract due to heterogeneity. Intein-mediated protein ligation and expression of a C1-C2 fusion protein in adenylyl cyclase deficient Escherichia coli were explored to circumvent the low affinity of the domains. However, the yields of products were insufficient for …
-
EXC-5 Controls Intracellular Trafficking in Order to Maintain the Apical Structure of the C. elegans Excretory Canal
… cell and in exc-5 mutants, the generation of intein containing proteins to create conditional alleles, and the generation of antibodies that bind to EXC-5. The data from these studies lead to a model of tubular maintenance where the sorting of material to the apical surface is only required at …
-
Studies on the Maturation of Secreted Quorum Sensing Peptides That Regulates S. Aureus Virulence
… these goals, an in vivo secretion assay using intein chemistry to produce AIP in the absence of AgrB was developed to examine whether AgrB facilitates AIP secretion. Also, SpsB biochemical assays were used to provide a thorough investigation of the final cleavage step in AIP biosynthesis for …
-
Engineering intracellular antibody libraries
The goal of this research is to understand how three different parameters affect single chain variable fragment (scFv) binding capacity. The parameters that were varied include the number of variable complementarity determining regions (CDRs), the number amino acids used to diversify CDRs, and …
-
Optimizing nuclear transgene expression and genome editing for high-value compound production in the green microalga Chlamydomonas reinhardtii
… recreated using the newly developed SpecR split intein tool for targeting two loci. Concluding all findings of this thesis, multiple novel tools were established for genetic engineering, including bicistronic expression, targeted integration, and development of a domesticated strain for elevated …
-
Structural and Biophysical Characterisation of Denatured States and Reversible Unfolding of Sensory Rhodopsin II
… protein complexes. Naturally occurring split inteins can mediate spontaneous trans-splicing both in vivo and in vitro. Previous studies have demonstrated successful assembly of proteorhodopsin from two separate fragments consisting of helices A–B and helices C–G via a splicing site in the BC …