Cloning and expression of human Eppin in E.coli
SHEN Zigang
CHEN Zhengqiong
HE Wei
YAN Ping
HE Haiyang
ZHANG Ji
YANG Xia
TANG Yan
WU Yuzhang
LIANG Zhiqing
LI Jintao
Abstract：Objective To construct prokaryotic expression vector for human Eppin gene and analyze the characteristics of expressed fusion protein.Methods Human Eppin cDNA was obtained from human testis tissue through RT-PCR,and then cloned into the pMAL-c2X vector.The recombinant protein was achieved in E.coli through IPTG induction.The expressed product was purified with Amylose resin,while the immunological activity of the expression product was detected by Western blotting.Results The gene fragment with length of 396 bp was amplified and coned into pMAL-c2X successfully.SDS-PAGE showed a protein band with relative molecular weight of 54 000,which was consistent with the expectation.The expressed product reached a high purity after Amylose resin chromatograph.Western blot method showed that the recombinant protein possessed good antigenicity.Conclusion pMAL-c2X-Eppin recombinant expression vector provides a base for studying of antifertility function of pMAL-c2X-Eppin recombinant protein.
Keywords：human EppinRT-PCRprokaryotic expressionrecombinant protein
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：4( 662-665 )