2004•Zhongguo xiandai yixue/Zhongguo xiandai yixue zazhiRequires access
Cloning, expression and identification of recombinant human trail
Xue-Gong Fan
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Abstract
Objective: To clone human Trail(TNF-related apoptosis-inducing ligand) gene into E. coli expression vector and to obtain recombinant human Trail (rhTrail) with biological activity.Methods:The outer-membrance region of human Trail cDNA was amplified from activated human peripheral blood lymphocytes using RT-PCR, and cloned into cloning vector pGEM-T. Then the clone of Trail cDNA gene was subcloned into the prokaryotic expressive pGEX-5X, followed by expression in E. coli HB 101 and purification. Results:The Results showed that the Trail cDNA was obtained and the pGEX-5X-Trail was also made, which can be expressed in E. coli. The apoptosis of HepG2 could be induced by the purified rhTrail. Conclusions:prokaryotic expression systems of human Trail gene with high efficiencies are successfully established. The expressed rhTrail with satisfied apoptosis-inducing activity can be useful for the further research.