The Construction and Expression of Recombinant Plasmid GFP-mip of Legionella Pneumophila Macrophage Infectivity Potentiator Gene

Objective To construct recombinant plasmid GFP-mip of Legionella pneumophila macrophage infectivity potentiator gene and observe its expression in the NIH3T3 cells.Methods The macrophage infectivity potentiator gene was amplified from DNA of Legionella pneumophila by polymerase chain reation(PCR),then cloned into pEGFP-C1 vector.The recombinant plasmid was named as GFP-mip and was analyzed with restriction endonuclease XhoI and BamHI digestion,PCR and DNA sequencing techniques.The NIH3T3 cell was transfected by recombinant plasmid GFP-mip with lipofection strategy.The stable expression products of macrophage infectivity potentiator gene were observed by the fluorescent microscope.Results 702bp mip gene was amplified.Under the fluorescent microscope,green fluorescent was observed in the cell cytoplasm and on the cell membrane.Conclusion The recombinant plasmid GFP-mip was constructed successfully and expressed in the NIH3T3 cells.