Participant: PROMISE AGEP Research Symposium
Department: Biological Sciences
Institution: University of Maryland, Baltimore County (UMBC)
A Molecular Screening Tool for the Detection and Quantification of Alteromonas macleodii
Alteromonas macleodii is a bacterium that displays a range of phenotypes which allows it to survive in many marine environments. Although A. macleodii has low representation in metagenomic datasets, it is able to contribute to the flux of organic material in marine environments. Furthermore, it is of interest to examine the copper tolerance mechanisms in A. macleodii to understand its ability to colonize copper-treated marine vessels. Currently, few tools exist that directly assess the general ecology of A. macleodii. To gain insight into this ecology, molecular screening tools are under development in our lab to: (1) determine the presence and (2) quantify the relative abundance of A. macleodii within marine ecosystems. An initial PCR screen was performed using universal primers specific to the 16S rRNA gene on DNA extracted from a variety of marine environments. After confirmation that extracted DNA was sufficient for amplification, primers targeting a 1,134 bp region of the DNA gyrase subunit B (gyrB) gene in A. macleodii were designed, optimized, and applied to the environmental samples. Reactions were visualized using gel electrophoresis, and samples yielding a band of the expected size were sequenced via Sanger sequencing to confirm specificity. To date, the A. macleodii-specific gyrB PCR assay has identified A. macleodii in a subset of the samples. Future work includes assuring primer specificity by testing the assay against multiple strains of A. macleodii, screening against additional marine environmental samples, and development of a specific quantitative PCR (qPCR) assay.
Translation in Saccharomyces cerevisiae
Translation is very efficient in Saccharomyces cerevisiae. In fact, translation codon errors vary from 4.0×10-5 to 6.9×10-4 per codon in this yeast species. It is hypothesized that the regulation of errors is controlled by Protein Kinase C 1 (Pkc1) via the help of a scaffolding protein Asc1. To observe the effects of Pkc1, a knockout of the Asc1 gene was performed in two strains: wild-type and ∆Ctk1. C-terminal Domain kinase 1 (Ctk1) phosphorylates the ribosomal protein Rps2 which increases translation efficiency. A decrease in translation efficiency has been observed when Ctk1 is depleted. The double knockout, ∆Ctk1∆Asc1, is hypothesized to have an increase in translation efficiency. After successful knockout of the Asc1 gene, the amount of errors will be quantified by a beta-galactosidase reporter gene system that carries a mutation at the 537 amino acid site. The frequency of errors can be calculated by comparing beta-galactosidase activity of the wild-type and mutant strains. Figuring out the details of protein synthesis will not only help to further the understanding of translation efficiency, but also the information gained can be used to make protein synthesis more efficient for biotech companies.
Brianda Beverley is a Biological Sciences Master’s degree student at the University of Maryland Baltimore County. She received her Bachelor’s degree in Biology from George Mason University in Fairfax, Virginia. Upon graduation, Brianda was accepted into and completed a Medical Laboratory Science certification program at INOVA Fairfax hospital. Her research interests include microorganisms involved in infectious diseases.
GENERAL SUMMARY OF GRADUATE RESEARCH
Alteromonas macleodii is a marine bacterium with a flexible genome that contributes to its wide range of phenotypes allowing it to survive in many marine environments. Our lab is specifically interested in the biofouling communities associated with copper-treated marine vessels and understanding the copper tolerance mechanisms employed by A. macleodii to colonize marine vessels. The primary goals of my research are to gain insight into the general ecology of A. macleodii in marine ecosystems by developing molecular screening tools that (1) determine the presence and (2) quantify the relative abundance of A. macleodii in marine environments.
SELECTED LIST OF PRESENTATIONS AND PUBLICATIONS
- November 15, 2018 – Annual Biomedical Research Conference for Minority Students (ABRCMS), Indianapolis, IN, “Developing Molecular Tools to Detect Alteromonas macleodii in the Environment” Brianda Beverley and Kathleen Cusick Ph.D. Poster Presentation
- October 31, 2018 – CAFPA-ASM DC Branch Fall 2018 Meeting FDA-CFSAN Harvey W. Wiley Federal Building, “A PCR Assay for Determining Alteromonas macleodii Presence and Diversity in Marine Environments” Brianda Beverley and Kathleen Cusick Ph.D. Poster Presentation
- March 28, 2018 – Graduate Research Conference University of Maryland-Baltimore County, Baltimore, MD “Translation efficiency in Saccharomyces cerevisiae” Brianda Beverley, Kartikeya Joshi, and Philip Farabaugh Ph.D. Poster Presentation
- February 16, 2018 – PROMISE AGEP Research Symposium University of Maryland, College Park, MD “Translation in Saccharomyces cerevisiae” Brianda Beverley, Kartikeya Joshi, and Philip Farabaugh Ph.D. Oral Presentation
Disclaimer: Information on this page has been provided by and is owned by the student presenter.