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Mutation of the gltT gene decreases the efficiency of growth of Neisseria gonorrhoeae

Kim, Daniel (2015)
Honors Thesis (35 pages)
Committee Chair / Thesis Adviser: Shafer, William M
Committee Members: Cafferty, Patrick William ; Campbell, Kathleen
Research Fields: Biology, Microbiology
Keywords: gonorrhea; glutamate transporter; cloning; gltT
Program: College Honors Program, Biology
Permanent url: http://pid.emory.edu/ark:/25593/pg27t

Abstract

The gltT gene encodes for an L-glutamate ABC transporter. In this work, I studied the effects of a gltT-negativemutation in streptomycin resistant Neisseria gonorrhoeae FA19 (FA19StR). A gltT-negative mutant, FA19StRgltT::kan, was shown to be deficient for growth in GC broth in comparison with its parental strain FA19. In the related pathogen N. meningitidis, acquisition of L-glutamate through GltT promotes evasion of polymorphonuclear leukocyte (PMN)-generated reactive oxygen species. Accordingly, oxidative stress due to hydrogen peroxide of FA19StRgltT::kan was tested, although the mutant FA19StRgltT::kan always appeared more sensitive to H2O2 than its isogenic parental strain FA19StR, the difference in sensitivity was not statistically significant. This work reveals genetic pathways that could potentially weaken gonococcal survival in PMN. Lastly, attempts to complement the gltT-negative mutant using the pGCC4 vector were unsuccessful. Sequence of the construct used for complementation (pGCC4gltT) revealed the presence of a new mutation in the coding region of the gltT gene that could result in the production of a non-functional protein.

Table of Contents

Introduction 1 -- Objectives 7 -- Methods 8 -- Results 11 -- Discussion 14 -- Future Research 17 -- Tables and Figures 18 -- References 25
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