Antibiotic and toxin resistance in commonly encountered pathogenic microorganisms is a growing worldwide healthcare issue. Although there is universal agreement about how antibiotic treatments function to inhibit bacterial growth, currently scientists are at a loss to explain how microorganisms are acquiring the ability to circumvent commonly implemented antimicrobial strategies. The looming crisis humans face as a result of wide-ranging antibiotic resistance drives the need for investigation into the mechanisms used by microbes to outpace our efforts. Recent studies have shown that the bacterial species Escherichia coli can become resistant to a wide range of antibiotics and cell toxins by simply overexpressing genes not typically associated with pathogenesis. One of the genes identified as a driver of antibiotic resistance from previous studies is ypjC (a largely uncharacterized membrane channel protein). This study seeks to determine the mechanism of antibiotic resistance conferred by overexpression of the gene ypjC in E. coli. Our initial line of investigation specifically targets the mechanism of action used by ypjC to confer resistance to ornidazole. In this investigation, we cloned ypjC using traditional polymerase chain reaction (PCR) methods; then, we inserted the amplified gene product into a cloning vector via digestion with restriction endonucleases and subsequent vector ligation. We followed this by transforming E. coli cells with the manufactured cloning vectors. Generation of this overexpression system is the first step in a larger project that ultimately seeks to investigate the impact of gene amplification on antibiotic resistance in a wide variety of genes found in E. coli.
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