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Full-Text Articles in Molecular Biology
Development Of A Genetic Over-Expression System For The Freshwater Sponge Ephydatia Muelleri, Joe Walsh
Development Of A Genetic Over-Expression System For The Freshwater Sponge Ephydatia Muelleri, Joe Walsh
Honors Theses
The aim of this work is to develop an over-expression system for Ephydatia muelleri. Preliminary work by Klaske Schippers (Schippers, 2013) showed sponges were induced to transcribe reporter genes driven by the widely used cytomegalovirus promoter (CMV). Building off Schippers work, newly identified native Ephydatia promoters were identified from the genome and verified to drive expression ofreporter genes. Validated over-expression vectors and transfection technology will allow researchers around the world who are working with this organism to ask questions about cell specification, gene regulation and animal development.
An Nmr-Guided Approach To The Isolation Of Secondary Metabolites From Nrps And Pks Gene Clusters, Connor Craig
An Nmr-Guided Approach To The Isolation Of Secondary Metabolites From Nrps And Pks Gene Clusters, Connor Craig
Honors Theses
Secondary metabolites continue to offer a promising source of novel chemical entities with therapeutic bioactivities. Our research aims to isolate secondary metabolites produced by non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) constructs. A metagenomic library was created from the marine bacterium Pseudoalteromonas sp., which was isolated from the Chesapeake Bay sponge Halichondria bowerbanki. Using a functional screen for PPTase activity we were able to identify colonies that contained NRPS and PKS gene clusters. Cultures of this microbe have been produced and analyzed using a combination of chromatography and 1H, 13C, HMBC, HSQC, COSY, AND NOESY NMR spectra. …
Isolation Of Microbial Nrps And Pks Gene Clusters For Natural Product Isolation, Danielle N. O'Hara
Isolation Of Microbial Nrps And Pks Gene Clusters For Natural Product Isolation, Danielle N. O'Hara
Honors Theses
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 …