Construction and Expression of Pip Gene of Legionella Pneumophila in Prokaryotic Cell

Objective To construct the recombinant plasmid pET-pip and to detect its expression in the prokaryotic cells BL21(DE3).Methods The pip gene was amplified by PCR from DNA of Legionella pneumophila,then inserted it into the prokaryotic expression vector pET-32a(+).The recombinant plasmid pET-pip was constructed.The recombinant plasmid was identified by restriction-endonuclease digestion,PCR and DNA sequencing analysis and transferred into BL21.The expression of pET-pip was induced with IPTG and the fusion protein was analyzed with SDS-PAGE.It was purified by His-tag.Results The pip gene of 726bp in length was amplified and the recombinant plasmid pET-pip was constructed.The PIP protein of approximately 46KD in size was expressed in E.coli and purified.Conclusion The pip gene of 726bp in length was successfully cloned and the recombinant plasmid pET-pip was constructed successfully and expressed efficiently.