Cloning, expression and identification of human survivin in E.coli

Abstract

Abstract: Objective To efficiently express and identify recombinant human survivin in E.coli. Methods Survivin cDNA was amplified by reverse transcriptional (RT)-PCR and cloned into the prokaryotic expression vector pBV220, followed by expression of the recombinant plasmid in E.coli strain BL21 (Gold). To obtain survivin protein, DEAE-Sepharose Fast-Flow ion exchange chromatography and Sephacryl S-200 gel filtration were performed. Western blot analysis was used for detecting the expressed product. Results Survivin protein was expressed in E.coli in the form of inclusion body at the expression level over 30% of the total cell protein. After ion exchange chromatography and gel filtration, the recombinant protein reached a purity over 95% and exhibited specific reaction with mouse anti-human antibody. Conclusion Survivin protein with high purity can be obtained by the method described above to facilitate further study of the anti-apoptosis function of survivin.

CLC Number:

BIE Jun¹, SUN Mao-sheng², SUN Qiang-ming², ZHENG Jian-ping³, SUN Wen-jia². Cloning, expression and identification of human survivin in E.coli[J]. Journal of Southern Medical University, 2004, 24(08): 913-916.

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