2010 Student Research Conference:
23rd Annual Student Research Conference

Comparison of Methods for Genetic Manipulation in Yersinia pestis: Deletion of the y0237 Gene
Jacob E. Schrum
Dr. Jon Goguen (University of Massachusetts Medical School) and Dr. Cynthia Cooper, Faculty Mentors

Yersinia pestis, the causative agent of plague, is a potential bioterrorism agent. The virulence of Y. pestis depends on the evasion of the lipopolysaccharide (LPS)Toll-like receptor 4 (TLR4) inflammatory response pathway. Low activity levels of LpxP are crucial for synthesis of LPS with low TLR4-stimulatory activity at human body temperatures. A gene of unknown function, y0237, is located immediately downstream of lpxP. It is found only in two Yersinia species and has little homology to any known gene; therefore, it is of interest to prepare an y0237 knockout mutant for future phenotypic analysis. Two methods were employed to delete y0237: a general method involving allelic exchange with a plasmid, and a recombineering method commonly used in Escherichia coli utilizing the recombination functions of bacteriophage λ, which requires fewer steps. The gene was successfully deleted using both methods, demonstrating the practicality of the recombineering method for routine use in Y. pestis.

Keywords: Yersinia pestis, microbiology, plague, infectious diseases, host-pathogen interactions, mechanisms of bacterial pathogensis

Topic(s):Biology

Presentation Type: Poster

Session: 8-4
Location: SUB-GEO
Time: 4:15

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