8th Annual WML Academic Symposium

Presenter(s): Nidhi Shah
Title: Utilization of ThyA genes from Geobacillus-infecting bacteriophages, GBK1 and GBK2 to develop a counter-selection system in an E. coli thyA – mutant strain.
Developed under the guidance of Mr. Timothy Marks , Pharmaceutical Sciences

Currently, antibiotic resistance markers are the standard to analyze and maintain vectors in biotechnology. But if the strains carry clinically significant antibiotic resistance genes, their use will be challenging. In this study, the goal is to develop a counterselection system and investigate thyA gene from Geobacillus-infecting bacteriophages, GBK1 and GBK2 as a potential gene marker in the E. coli thyA mutant strain without relying on antibiotics. The two novel thyA genes will be isolated from both bacteriophages using PCR and each will be ligated into the pJZ04e vector. The recombinant plasmids will be transformed into the mutant strain and will be grown in media lacking thymine. The recombinants will be confirmed by performing PCR.