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.
Recommended Citation
O'Hara, Danielle N., "Isolation of microbial NRPS and PKS gene clusters for natural product isolation" (2015). Honors Theses. 908.
https://scholarship.richmond.edu/honors-theses/908