A Mutational Analysis of Bacteriophage Lambda Site-Specific Recombination (In Vitro, Dna-Dna Interaction, Protein, Oligonucleotide-Directed)
Bauer, Carl Eugene
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https://hdl.handle.net/2142/71176
Description
Title
A Mutational Analysis of Bacteriophage Lambda Site-Specific Recombination (In Vitro, Dna-Dna Interaction, Protein, Oligonucleotide-Directed)
Author(s)
Bauer, Carl Eugene
Issue Date
1986
Department of Study
Microbiology
Discipline
Microbiology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Molecular
Biology, Microbiology
Abstract
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 dependent DNA-DNA interactions only occur within the 7-base "overlap" region where strand exchange occurs. In vitro recombination analyses with mutant att sites further demonstrated that the homology dependence involves DNA interactions between one strand in each recombining att site. The in vitro analyses also demonstrated that homology dependent interactions appear to occur at both the synaptic and strand resolution stages of recombination. The segregation patterns of single point mutations within the core were also followed from the parent into the recombinant att sites. The mutations segregated at a predicted pattern that was dependent on the location of the mutations relative to the position of strand exchange. A related set of point mutations were also constructed within each of the five Int arm-type binding domains. Footprint analysis and recombination analysis of individual and collective mutations demonstrated that Int interacts with unique arm-type sites during integrative and excisive recombination.
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