Cloning, expression, and identification of the endolysin encoded by bacteriophage MmP1 against Morganella morganii
ZHU Xiaoyan
ZHU Junming
WANG Jing
LI Shu
RAO Xiancai
HU Fuquan
TAN Yingling
Abstract：Objective To construct,express,and purify the recombinant endolysin encoded by Morganella morganii bacteriophage MmP1 for the detection of its biological function.Methods The objective endolysin gene was amplified from the genome of bacteriophage MmP1,and then cloned into a procaryotic expression vector to construct recombinant plasmid pQE31-M12,which was subsequcntly transformed into E.coli JM109.The affinity chromatograph plus gel chromatography was applied to purify the endolysin protein under native conditions.Antibacterial activity of the endolysin was detected by gel diffusion method.Results The sequence of recombinant plasmid pQE31-M12 was identified to be correct through enzyme digestion and sequencing.The secrete expression of recombinant endolysin from engineering bacteria could be induced by 0.5 mmol/L IPTG for 5 h at 37℃.The endolysin protein was purified from supernatant of ultrasonic-treated bacteria by nickel-affinity chromatography using gravity-flow.and desalted on a molecular-sieve chromatography.The growth of S.aureaus could be inhibited obviously by endolysin.Conclusion The recombinant E.coli containing endolysin gene has been successfully constructed and the purified endolysin protein with antibacteria activity against S.aureaus has been obtained,which lay a foundation for the reshuffling of endolysin structure and enhancing its biological activities.
Keywords：bacteriophageendolysingene expressionpurification
Publication Date：2009-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 705-708,712 )