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 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.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Microbiology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Bauer, Carl Eugene

• Biology, Molecular
• Biology, Microbiology

Identifier

(UMI)AAI8610905

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/71176