Exploring the role of OmpC deletion and O-antigen expression in T4 bacteriophage-induced lysis of Escherichia coli K12 cells

Abstract

Escherichia virus T4 (T4 phage) is a species of bacteriophage that infects Escherichia coli (E. coli) bacteria. Resistance to T4 phage infection has been found in E. coli strains that express the outermost component of lipopolysaccharide, O-antigen, or in those that do not exhibit outer membrane porin C (OmpC). However, the exact mechanism by which resistance is conferred in both cases has yet to be elucidated. E. coli K12 strains lack O-antigen and are susceptible to T4 phage infection due to a mutation in wbbL, a rhamnosyltransferase gene necessary for O-antigen synthesis. As such, we sought to explore the combined effect of ompC deletion and O-antigen expression in T4 bacteriophage-induced lysis by creating a plasmid that contains the wbbL gene, followed by its introduction into an ompC knock-out strain. Using molecular cloning techniques, an intact copy of wbbL was inserted into a plasmid vector as confirmed by restriction enzyme digestion, gel electrophoresis, and Sanger sequencing. The plasmid, designated pCR2.1-wbbL-