2010 Summer NC State Undergraduate Research Symposium

Session Time : 8/5/10 1:00 PM - 8/5/10 4:00 PM

Content Area : HHMI Reaching Incoming Students Enrichment (RISE)

Student Presenters :
Kaitlyn Eileen Casulli Food Science

Mentors and/or Co-Authors :
Sophia Kathariou Food, Bioprocessing & Nutrition Sciences

Abstract Title : Vector construction for generation of kpsM mutants in multi-drug resistant Campylobacter strains

Abstract :
Campylobacter is a major foodborne bacterial pathogen primarily transmitted through poultry. Capsular polysaccharides are important for various adaptations in bacteria, and have been shown to be critical for pathogenesis in Campylobacter. However, limited understanding is available on the role of capsule in colonization of avian hosts. The primary objective of this project is to generate a cloning vector with a chloramphenicol resistance marker inserted into the capsule biosynthesis kpsM gene of Campylobacter, thus disrupting the gene and resulting in absence of capsule. The kpsM fragment came from a turkey-derived strain of C. coli (6067) from our lab collection and has recently been sequenced. This kpsM gene was inserted into pUC19, which is commonly used as a cloning vector for Escherichia coli, and then the chloramphenicol resistance cassette derived from pJMA-001 was inserted into kpsM. pUC19 was chosen for this project because of its ampicillin resistance and blue/white screening capability. For Campylobacter, kpsM is important for proper capsule formation. The final plasmid construct will be used to create kpsM mutants of multi-drug resistant Campylobacter strains which will be used in future phenotypic assays.