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Controlled expression of lysis gene E by a mutant of the promoter pL of the thermo‐inducible λcI857‐pL system
Author(s) -
Fu L.X.,
Gong J.S.,
Gao B.,
Ji D.J.,
Han X.G.,
Zeng L.B.
Publication year - 2021
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.14690
Subject(s) - lytic cycle , lysis , mutant , mutagenesis , escherichia coli , biology , microbiology and biotechnology , plasmid , gene , mutation , lambda phage , chemistry , biochemistry , genetics , bacteriophage , virus
Aims To identify a lambda promoter pL mutant that could extend the thermal stability of the thermo‐inducible λcI857‐pR/pL system and to evaluate the effects of the modified system for the controlled expression of lysis gene E during the production of bacterial ghosts (BGs). Methods and results The promoter pL mutant was identified by random mutagenesis and site‐directed mutagenesis. The results showed that a T → 35C mutation in the pL promoter was responsible for the phenotype alteration. Under the same induction conditions, the lysis rates of the modified lytic system on Escherichia coli and Salmonella enteritidis were significantly lower than that of the control, while the lysis rates of Escherichia coli with the thermo‐inducible lytic system were significantly higher than that of S. enteritidis with the corresponding plasmid ( P < 0·05). Conclusions Increasing the heat stability of the thermo‐inducible lytic systems decreased lysis efficiency during the production of BGs. There exist differences in the lysis efficiency of thermo‐inducible lytic systems between different bacterial strains. Significance and Impact of the Study These findings enrich current knowledge about modifications to thermo‐inducible systems and provide a reference for the application of these modified systems for the production of BGs and controlled gene expression in bacteria.