Determining Genetic Causes of Streptococcus pyogenes' Susceptibility to Retinoic Acid

Overview

Objectives/Goals This project's goal was to find the genes that contribute to Streptococcus Pyogenes' susceptibility to retinoic acid, the active form of Vitamin A. Methods/Materials A genetic screen was used to find the retinoic acid resistant AP M1T1 GAS 5448 mutants created using a Transposon Tn917 library. This was done through screening the transposon library, alternating it in 5µg/mL erythromycin antibiotic in Todd-Hewitt Broth (THB) and increasing concentrations of retinoic acid in 80% Roswell Park Memorial Institute media and 20% THB. After a manageable number of mutants were found, their genes disrupted by the transposon insertions were discovered using single-primer polymerase chain reactions, GeneWiz sequencing services, and ncbi.gov's nucleotide BLAST tool. Results To date, three genes have been identified that contribute to susceptibility to retinoic acid; CitG gene, Csn1 gene and UvrC from the mutants A9 Sequence L-2, B6 Sequence L-2 and B10 Sequence L-2, and D10 Sequence L-2 respectively. Conclusions/Discussion The CitG gene either codes for part of the GntR family of transcriptional regulators, or a catalyst of functions dealing with energy as ATP. The Csn1 gene codes for proteins associated with prokaryotic acquired immunity. The UvrC gene codes for 1 of 3 subunits of an endonuclease that participates in the nucleotide excision repair process. These genes and how they contribute to the bacterium#s susceptibility can be carefully studied further in the future to create novel non-antibiotic treatments of Streptococcus infections, thus reducing society#s dependence on antibiotics and slowing the proliferation of antibiotic-resistant bacteria.

Awards (1)

• Fourth Place

Competition history

• CSEF 2015 Microbiology (General) · Entry S1504

Resources

• CSEF project summary ↗

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