Construction of Recombinant Plasmid and Fusion Expression of rhIL-10 in BL21(DE3)pLysE Cells

Objective To construct the recombinant plasmid and express the fusion protein of rhIL 10 in E.coli. Methods Amplify IL 10 gene by RT PCR,clone the PCR product to construct recombinant plasmid PCRT7/NT TOPO IL 10,and identify by Appied Biosystems 3 700 DNA analyzer.Transform the constructed plasmid into E.coli BL21(DE3)pLysE cells and identify the expressed fusion protein by 12% SDS PAGE.Results rhIL 10 gene was successfully inserted into vector PCR T7/NT TOPO.The sequence of the insert was completely consistent with that desinged.The insert in the recombinant plasmid was highly expressed mainly in inclusion bodies in BL21(DE3)pLysE cells.Conclusion Recombinant plasmid PCR T7/NT TOPO IL 10 was successfully constructed and IL 10 fusion proteins highly expressed in BL21(DE3)pLysE cells.