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.
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Showing 1 to 20 of 21 for “"Site-specific recombination"”.
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Protein/DNA interactions during site-specific recombination
… that IntDOT interacts with five arm-type sites, but exactly how the arm-type sites participate in the integration and excision of CTnDOT was not known. I constructed site-directed mutations in each of the arm-type sites and tested them in in vitro integration and excision assays. In vitro …
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Coordination and Catalysis of the Lambda Integrase Site -Specific Recombination Reaction
In the course of my doctoral work, I have developed quantitative assays to measure the maximal cleavage and ligation activity of Integrase. From this work, we concluded that para-Nitrophenol tyrosine analogs are optimal for quantitative in vitro measurement of Int ligation. Int could not utilize …
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The P22 Xis Protein: Regulation of Bacteriophage P22 Site -Specific Recombination
… DNA with a weaker affinity but still formed specific complexes.
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Analysis of Structure-Function Relationships in *Lambda and HK022 Site -Specific Recombination
… Lambda and HK022 Int core-type DNA binding specificity was examined by employing challenge phages to isolate gain-of-function HK022 Int mutants that recognize the Lambda B' core site. The HK-Int D99 residue was identified as a negative specificity determinant that apparently contacts the …
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An Analysis of Bacteriophage Lambda Site-Specific Recombination Using IHF Binding Site Mutants
… of bacteriophage lambda. Mutations of the three sites to which this protein binds within the lambda attachment site were constructed using oligonucleotide-directed site-specific mutagenesis and were ananlysed for their effects on site-specific recombination. One group of mutations had alterations …
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Mutational analysis of integrase and excisionase proteins in bacteriophage lambda site-specific recombination system
Second-site reversion analyses were performed on 21 recombination defective Int mutants. A second-site revertant P243L/E218K and a pseudorevertant E175K were isolated and further characterized using genetic and biochemical methods. The results showed that the second-site substitution E218K …
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A Mutational Analysis of Bacteriophage Lambda Site-Specific Recombination (In Vitro, Dna-Dna Interaction, Protein, Oligonucleotide-Directed)
Mutations within the lambda attachment (att) site were constructed by oligonucleotide-directed site-specific mutagenesis and analysed for their effects on site-specific recombination. The phenotype of mutations within the 15 base "core" region of sequence homology demonstrated that homology …
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Genetic analysis of the proteins required for site-specific recombination of bacteriophage lambda: Interactions with each other and their recognition sites
… of the individual Int arm-type binding sites in $\lambda$ site-specific recombination, triple-base changes were made in each of the five Int arm-type binding sites, and then characterized through in vitro and in vivo recombination assays. These mutants demonstrated that the P1, …
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Evaluation of the Recombination Efficiencies of FLP Proteins
<p>Site-specific recombination systems are powerful tools for genetic modification. They have been used to integrate a transgene into a pre-defined locus and to remove marker genes from a transgene locus. Two of the most widely used site-specific recombination systems in plants are the Cre/lox …
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A genetic analysis of bacteriophage lambda integrase-core site interactions
Bacteriophage lambda utilizes a site-specific recombination system to integrate its DNA into the E. coli chromosome during lysogeny. The viral protein, Integrase (Int), catalyzes strand exchange between the phage attachment site, attP, and the host target sequence, attB to generate the prophage …
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Bacteroides Conjugative Transposons: Ecology and Mechanisms
… Previous studies had shown that the attDOT recombination site and integrase of CTnDOT (IntDOT) are sufficient and essential for integration in vivo. Even though the integrase of CTnDOT belongs to the lambda integrase family of recombinases, the mechanism of integration of CTnDOT is different …
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Analysis of Spacer Region derived from Recombination between Heterologous loxPs
"목적 : 부위특이적 재조합 (site-specific recombination) 방법의 하나인 Cre-loxP 시스템에서, mutant loxP 간의 재조합 반응 시 spacer region 염기 서열을 분석한다. 또한 heterologous (mutant) loxP 간의 반응 시 spacer region의 염기 서열 분석을 통하여 Cre 매개 재조합 메카니즘의 이해를 넓힌다. 재료 및 방법 : 두 lox site 사이의 Cre 매개 재조합 반응으로 나타나는 spacer region의 염기 서열 분석을 위하여 새로운 벡터 …
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Genetic and mechanistic studies of the bacteriophage lambda integrase protein
"Site-specific recombination between bacteriophage $\lambda$ and E. coli chromosomes starts with the formation of higher-order nucleoprotein complexes, called ""intasomes"", followed by a series of steps catalyzed by the phage-encoded integrase (Int). In order to study the functions of Int during …
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A study of DNA-DNA interactions during bacteriophage lambda integrative recombination
An in vitro integrative recombination system was used to study DNA-DNA interactions during site-specific recombination in bacteriophage lambda. Oligodeoxynucleotides 28 bases long containing either the top strand or the bottom strand sequences of the bacterial attachment site (attB) with or without …
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The Engineering of Chinese Hamster Ovary Cells to Achieve More Efficient Gene Amplification for Improving Biopharmaceutical Development
… of therapeutic proteins. It addresses the specific time bottlenecks associated with developing high producing CHO cell lines and reviews various methods that are employed to alleviate these bottlenecks. The specific time consuming process of gene amplification is the focus of this work. …
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Mathematical and experimental approaches to the dimer catastrophe theory
… host. Plasmid dimers emerge through homologous recombination. Dimers have two independent origins of replication and thus have a replicative advantage and reduced copy number. Models of plasmid behaviour suggest that dimers would overtake a cell population, but that this can be prevented if they …
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Improving Bacterial Therapeutics and Diagnostics through Synthetic Biology
… concurrent efforts to enhance tunability, specificity, efficacy, and safety. Our work contributes to three such areas. First, we performed model-guided engineering of DNA sequences with predictable site-specific recombination (SSR) rates, demonstrating that recombinase attachment sites with …
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Investigating the role of IL-4/IL-13 signalling through the IL-4 receptor alpha (IL-4Rα) on keratinocytes in murine models of Leishmania major and Schistosoma mansoni
… the skin is important during the parasite life cycle. While Th1 immunity is required to control CL, protection during schistosomiasis requires Th2 immunity. Paradoxically, Th2 characteristic IL-4 secreted early during L. major infection in mice, can drive a Th1 response by instructing …
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Investigating R gene evolution by meiotic recombination using synthetic gene clusters in Arabidopsis
… (synthRPP1) to model R gene evolution by meiotic recombination. This experimental design utilizes gain-of-luciferase phenotype (luc+) to identify and isolate recombinant R genes and uses two alternatively marked alleles to distinguish and measure different types of meiotic recombination (intra- …
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Development of broadly applicable transgenic tools for the transposon mutagenesis of the red flour beetle, Tribolium castaneum
The discovery of non-species-specific, broad-range transposable elements and the establishment of a universal 3xP3 promoter revolutionized insect transgenesis. It overcame the limitations of the germ-line transformation to be restricted to the model organism Drosophila melanogaster. In combination …
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