Use of cloned T4-phage gene to disrupt escherichia coli

摘要

Three lysis genes, gene-5, gene-t and gene-e of bacteriophage T4 were cloned in pUC118 and pET26b(+) vectors, respectively. Expression of gene product (gp) -t in the E. coli cells leaded prompt cell lysis. On the other hand, productionso f gp-5 and gp-e in the cytosol did not lead cell lysis. Translocation of the gp-e to the periplasmic space driven by signal peptide of pelB outer-membrane protein of E.coli fused in fram at N-terminal end of gp-e caused morphological change of the host cell. Rod shaped E. coli cells changed to the elliptical form. Resuspension of this gp-e producing cells with pure water caused cell lysis followed by beta-galactosidase release to the medium.