Date of Award

2015

Document Type

Thesis

Degree Name

Bachelor of Science

Department

Biochemistry & Molecular Biol.

First Advisor

Dr. Jonathan Dattelbaum

Second Advisor

Dr. Ellis Bell

Abstract

Greater than 99% of microbes living in the environment are uncultivable due to their complex nutrient and temperature requirements for growth. These microorganisms present a potential source of natural products that could be developed for biotechnological and pharmaceutical uses. Microorganisms with phosphopantetheinyl transferase (PPTase) activity are of high interest due to the role PPTase plays in activating non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) gene clusters. These gene clusters produce multi-enzymatic, multi-domain megasynthases that produce complex natural products utilized by the host organism for selective advantages. This study isolated genomic DNA from soil samples collected from the University of Richmond campus to construct metagenomic libraries with a host organism containing a BpsA reporter plasmid. These libraries were then screened for functional activity of NRPS and PKS gene clusters by virtue of PPTase activity. We then extended this study to include a library of pigmented microbes previously isolated from Chesapeake Bay sponges, Clathria prolifera and Halichondria bowerbanki. Genomic analysis of positive clones has thus far been inconclusive, however putative NRPS gene clusters have been identified in the SW202 microbial genome.

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