Using error prone PCR in directed evolution to selected novel antibiotic resistances

Abstract

The evolution of antibiotic resistance presents an escalating problem in the treatment of various infectious diseases worldwide. Although the origin of antibiotic resistance genes is not generally clearly documented, it has been thought that they evolved from specific genetic elements which eventually managed to spread to other microorganism of different strains and species through mobile genetics elements, transposons and plasmids. Extensively studying all aspects of these genes and their impact on the development of new treatments and drugs is of extreme importance. This study focuses on evolving and understanding how novel antibiotic resistance develops. Error prone PCR (EP-PCR) was used to introduce random mutation in an arr gene which confers high level resistance to rifampicin in E. coli. The clones obtained from EP-PCR were screened on different antibiotics with varying concentration in an attempt to isolate a clone with an increased minimum inhibitory concentration (MIC) as compared to the wild type parent strain (pBstN49).

Several clones showed decreased levels of resistance against rifampicin but however none showed any significant increase in any of the other antibiotic MICs tested.