An Analysis of Bacteriophage Lambda Site-Specific Recombination Using IHF Binding Site Mutants

Integration Host Factor (IHF) is an Escherichia coli protein required for integration and excision 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 across one site; in a second group, a pattern of changes was made in each of the three sites. Certain base changes affected some sites more severely than others; some changes had more effect in vitro than in vivo. All mutations depress integrative recombination in vivo and depress or eliminate it in vitro. In excision, mutations of the H1 site raise the frequency of excisive recombination, while mutations of the H2 or H$\sp\prime$ sites lower the frequency of excisive recombination. These results indicate an involvement of all three sites in both integration and excision; occupancy by IHF of the H2 and H$\sp\prime$ sites is required for either reaction, while the presence of IHF at H1 favors integration and its absence favors excision.

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

• Waechter-Brulla, Daryle A.

• Biology, General

Identifier

(UMI)AAI8908882

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/72521