2004•Zhongguo renshougonghuanbing zazhiRequires access
Cloning and expression of the mip gene of Legionella pneumophila
Jian Chen
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Abstract
To construct the recombinant plasmid pLpmip and to detect its expression in the prokaryotic cells JM109,the mip gene of Legionella pneumophila was amplified by PCR from a template of sero group I of this organism.The amplified DNA was digested with BamH 1 and Sal 1 and ligated into vector pUC18. The recombinant plasmid was identified by restriction analysis and PCR with further confirmation by sequencing analysis.SDS PAGE and Western blotting were used to detect the expression of mip gene products.It was found that mip gene of 828 bp in length was amplified,and the recombinant plasmid could be constructed. The protein expressed in the prokaryotic expression system had a size of 24 kDa. It concludes that the mip gene of L pneumophila is successfully cloned and expressed in the present study.