Abstract: In order to find new drugs combined with the sensitive targets to solve the problem of drug resistance of pathogenic bacteria, by constructing the overexpression strain of the outer membrane protein ompC of Escherichia coli, the killing effect of gentamicin on it was tested and compared with that of the wild-type Escherichia coli. Then, the relationship between the protein expression level of ompC and the killing effect of gentamicin was explored by using the immunoblot assay, and the over-expression strain MIC was further determined. The results showed that the overexpression of ompC could significantly enhance the sensitivity of Escherichia coli to gentamicin, and after the overexpression of ompC, the minimum inhibitory concentration of the strain W3110-pCA24N (ompC) was 8 ug·mL^-1, which was significantly lower than that of the strain W3110-pCA24N (wild type) (being 16 ug·mL^-1). The gentamicin was more intaken into the cells of Escherichia coli, which enhanced the bactericidal effect of gentamicin. Therefore, the outer membrane protein was expected to be a new candidate target of drug sensitivity.