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Articles 1 - 4 of 4

Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Identification Of Bacterial Species Using Colony Pcr, Kaiti Walker Apr 2015

Identification Of Bacterial Species Using Colony Pcr, Kaiti Walker

Honors Theses

My research consisted of two separate parts, both involving the PCR methods. In part one I was trying to identify bacteria isolated from a chicken coop. In part two I was trying to identify bacteria isolated from the bluff. In both parts, I used a technique called colony PCR. This concept uses a single colony of bacteria to complete the PCR reaction. One pipet tip from a single colony is selected from the plate and mixed into a tube containing ultrapure water. A small amount of this bacterial mixture is then added into the PCR reaction. In the first part ...


Development Of A Genetic Over-Expression System For The Freshwater Sponge Ephydatia Muelleri, Joe Walsh Jan 2015

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 Jan 2015

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. We were ...


Isolation Of Microbial Nrps And Pks Gene Clusters For Natural Product Isolation, Danielle N. O'Hara Jan 2015

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 ...