title: Enhancement of bacteriolysis of shuffled phage PhiX174 gene E.
abstract: Bacterial ghosts that are generated using the regulated PhiX174 lysis gene E offer a new avenue for the study of inactivated vaccines. Here, we constructed a library of mutant gene E using a gene-shuffling technique. After screening and recombination with the prokaryotic non-fusion expression vector pBV220, two lysis plasmids were selected. Among which, a novel mutant E gene (named mE), consisting of a 74-bp non-encoding sequence at 5'-end and a 201-bp gene ΔE, significantly increased the lysis effect on prokaryotic Escherichia coli and Salmonella enteritidis. Moreover, lysis efficiency, as measured by the OD600 value, reached 1.0 (10⁹ CFU), avoiding the bottleneck problem observed with other bacterial lysis procedures, which results in a low concentration of bacteria in suspension, and consequent low production of bacterial ghosts. Our results may provide a promising avenue for the development of bacterial ghost vaccines.
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Science, Harbin 150001, China.
authors.forename: Shen-ye
authors.initials: SY
authors.lastname: Yu
authors.forename: Wei
authors.initials: W
authors.lastname: Peng
authors.forename: Wei
authors.initials: W
authors.lastname: Si
authors.forename: Lu
authors.initials: L
authors.lastname: Yin
authors.forename: Si-guo
authors.initials: SG
authors.lastname: Liu
authors.forename: Hui-fang
authors.initials: HF
authors.lastname: Liu
authors.forename: Hai-ling
authors.initials: HL
authors.lastname: Zhao
authors.forename: Chun-lai
authors.initials: CL
authors.lastname: Wang
authors.forename: Yue-hong
authors.initials: YH
authors.lastname: Chang
authors.forename: Yue-zhi
authors.initials: YZ
authors.lastname: Lin
mesh_terms: D001433:Bacteriolysis; D010584:Bacteriophage phi X 174 / Q000235:genetics* / Q000254:growth & development*; D032621:DNA Shuffling; D004279:DNA, Viral / Q000235:genetics*; D004926:Escherichia coli / Q000821:virology*; D010957:Plasmids; D012477:Salmonella enteritidis / Q000821:virology*
chemical_list: D004279:DNA, Viral