Cloning, expression and purification of gene products involved in peptidoglycan synthesis for Chlamydia trachomatis Public Deposited

http://ir.library.oregonstate.edu/concern/undergraduate_thesis_or_projects/wh246x73t

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  • Chlamydiae are obligate intracellular bacterial pathogens with a unique biphasic life cycle that consists of an infectious but non-dividing form, called elementary bodies (EB), and a noninfectious but metabolically active form, called reticulate bodies (RB). Chlamydia has an unusual process of cell division of which still remains uncharacterized subject matter; in particular, the paradox of peptidoglycan (PG) synthesis. A paradox is a principle or proposition contrary to received opinion (18). Chlamydaal research has shown evidence in support of and against PG synthesis. The intention of this project incorporates clarification of the role of PG in Chlamydia, which would result in a greater understanding of chlamydaal cell division. The primary objective of my research was to clone, express, and purify the three genes targeted for PG synthesis in C. trachomatis: pbp3, ftsW, and p60. This project incorporates many molecular biology techniques such as designing primers, PCR, restriction digests, transformation, PCR screening, cell culture, sequencing, column purification, and Western blot analysis. Unfortunately, this project was weighed down with much experimental difficulty, which produced subjective results.
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