Title page for E-project-042607-162955

Project Type MQP

Submission date 2007-04-26

Author
Audrey N Jajosky, BIO

URN E-project-042607-162955

Sponsor Dartmouth Medical School

Title Minimal Requirements for Assembly of a Stable Inner Membrane Platform for Vibrio cholerae Pilus Biogenesis

Advisor
Rulfs, Jill, BB

Availability unrestricted

Abstract
V. cholerae colonizes the human intestine via TCP & can cause diarrhea with fluid losses severe enough to be fatal. Without TCP, V. cholerae cannot attach to the intestine, secrete toxins, or cause GI disease. 4 proteins required for TCP biogenesis -- TcpD,E,R,& T -- localize to the IM. Previous experiments support the hypothesis that interactions among 3 of these 4 proteins are predicted to be sufficient to form a stable IM platform in V. cholerae. Plasmids were engineered to carry combinations of tcp genes and induced to co-express the 4 IM Tcp proteins in all groups of 2 & 3 in E. coli. Protein immunoblots on cell lysates suggest that TcpD and TcpE stabilize the TcpR-TcpT interaction. Therefore, 3 proteins required for TCP biogenesis appear to form a stable IM platform.

Files
07D367M.pdf

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Project Type	MQP
Submission date	2007-04-26
Author	Audrey N Jajosky, BIO
URN	E-project-042607-162955
Sponsor	Dartmouth Medical School
Title	Minimal Requirements for Assembly of a Stable Inner Membrane Platform for Vibrio cholerae Pilus Biogenesis
Advisor	Rulfs, Jill, BB
Availability	unrestricted
Abstract V. cholerae colonizes the human intestine via TCP & can cause diarrhea with fluid losses severe enough to be fatal. Without TCP, V. cholerae cannot attach to the intestine, secrete toxins, or cause GI disease. 4 proteins required for TCP biogenesis -- TcpD,E,R,& T -- localize to the IM. Previous experiments support the hypothesis that interactions among 3 of these 4 proteins are predicted to be sufficient to form a stable IM platform in V. cholerae. Plasmids were engineered to carry combinations of tcp genes and induced to co-express the 4 IM Tcp proteins in all groups of 2 & 3 in E. coli. Protein immunoblots on cell lysates suggest that TcpD and TcpE stabilize the TcpR-TcpT interaction. Therefore, 3 proteins required for TCP biogenesis appear to form a stable IM platform.
Files	07D367M.pdf