EXPRESSION AND FUNCTION OF CONSERVED BACTERIAL PROTEINS

University Of Md Biotechnology Institute, Baltimore MD

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Abstract

DESCRIPTION: The overall objective of this proposal is to define the function of evolutionary conserve genes of unknown function. Each gene of interest will be cloned into an intein vector to facilitate expression and purification. Cloned proteins that are expressed at high level in E. coli will be purified by chitin affinity chromatography followed by intein hydrolysis. The PI anticipates producing purified proteins in assembly line fashion with an output of approximately 20 purified proteins per year. These purified proteins would be provided to collaborators for three-dimensional structure determinations by X-ray crystallography and NMR spectroscopy. In addition, gene disruptions will be created, and the phenotypes of the resulting mutants will be determined under a variety of growth conditions.