Genetic characterization and investigation of kanamycin susceptibility of ompC and ompF single gene deletion mutants of Escherichia coli K-12

09/09/2018

Chuan Chuan Chang, Ching Wing Yan, Regina Onal, and Yue Zhang​

Volume 22
Fall 2017 / Winter 2018

SUMMARY OmpF and OmpC are proteins that form porins in the outer membranes of Gram-negative bacteria. Though the two porins have been shown to transport a variety of antibiotics, much remains unknown about aminoglycoside transport through OmpF and OmpC porins as previous studies relating OmpF to kanamycin resistance have shown inconsistent observations. We first performed genetic characterization of kanamycin sensitive Escherichia coli K-12 ompC and ompF single gene deletion mutants, which were generated from the removal of a kanamycin resistance cassette through FLP-FRT recombination of the original Keio collection ΔompC and ΔompF mutants. We then performed minimum inhibitory concentration assays to determine the lowest inhibitory kanamycin concentration of the mutants. We have shown the gene deletion mutations are accurate and precise in our Keio mutants, and we have preliminary results showing that the ompF deletion mutant has a greater resistance to kanamycin in comparison to the wildtype and ompC deletion mutant. This finding implicates that ompF may play a greater role in kanamycin susceptibility.