The expression,purification and preliminary use of human Survivin protein
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    Abstract:

    Objective:To clone the cDNA of human Survivin,study its expression in E. Coli. and demonstrat its significance in the diagnosis of hepatoma. Methods:Using the isolated total RNA from human leukemic cell line HL60 as a template,the human Survivin encoding cDNA was amplified by reverse transcription-polymerase chain reaction(RT-PCR) methods. The PCR product was cloned into prokaryotic expression vector pMD-18T and sequenced. The expression of recombinant plasmid pGEX-2TK-Survivin was induced in E.coli BL21 by IPTG, the expressed product was purified by GST protein purification system,then was cut GST from the expressed product useing Thrombin. After the identification of the Survivin protein by SDS-PAGE and Western blot,they were coated in ELISA plate to detect the expression in serum of hepatoma patient. Results:Human Survivin,an anti-apoptosis gene,express Surviving protein with a relative molecular mass of 16.2 ku was cloned and used preliminary in hepatoma patients. Conclusion:Human Survivin gene was cloned successfully;Survivin protein was successfully expressed,purified and preliminary used. It may provide preliminary labratory evidence based on Survivin for the diagnosis and therapy of carcinoma.

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李 烜,梁文飚,孙倍成,吴 华.人Survivin蛋白的表达?纯化及初步应用[J].南京医科大学学报(自然科学版英文版),2008,28(10):1267-.

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  • Received:May 22,2008
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