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Cloning, High Level Expression and Purification of Human Survivin |
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Abstract Survivin is a protein that inhibits apoptosis and regulates cell division. In our study, The cDNA sequence of survivin was amplified by RT-PCR and sub-cloned into the prokaryotic expression vector pET-21b(+), followed by transformation into E.coli strain BL21(DE3) and induction with IPTG. The recombinant survivin protein fusing with 6xHis tag was expressed in E.coli in the form of inclusion body at the expression level over 60% of the total cell protein. Results of Western blotting showed that recombinant survivin reacted specifically with anti-human survivin antibody. After gel filtration, the recombinant protein reached the purity over 95%, which facilitate the study of diagnosing and inhibitor agents targeting survivin.
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Received: 26 May 2007
Published: 25 October 2007
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