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Preliminary Research on the Expression, Purification and Function of the Apoptotic Fusion Protein, Sival

  • Zhang, Ya-Han (Suzhou Industrial Park Center for Disease Control and Prevention) ;
  • Yu, Lu-Gang (Suzhou Industrial Park Center for Disease Control and Prevention) ;
  • Zhu, Wan-Zhan (Suzhou Industrial Park Center for Disease Control and Prevention) ;
  • Wang, Sheng-Li (Suzhou Industrial Park Center for Disease Control and Prevention) ;
  • Wang, Dian-Dong (College of Life Science and Technology, Yangtze Normal University, Chongqing) ;
  • Yang, Yan-Xin (CSPC Zhongqi Pharmaceutical Technology) Co., LTD.) ;
  • Yu, Xia (Suzhou Center For Disease Prevention And Control)
  • Published : 2014.11.06

Abstract

The objective of the present study was to investigate cloning, expression, and functions of the recombinant protein, Siva1. Siva1 gene was synthesized by RT-PCR from HCT116 cells. Plasmids were cleaved with the restriction endonuclease, BamH1/Sal1 and products were connected to pQE30, which underwent cleavage by BamH1/Sal1. The recombinant plasmid, pQE30-Siva1, was identified after digestion with restriction endonucleases followed by transformation into E. coli M15. Expression of Siva1 was induced by IPTG and identified by SDS-PAGE following purification with affinity chromatography. The results showed that size of Siva1 was 12 kDa, consistent with the molecular weight of the His-Siva1 fusion protein. Functional test demonstrated that Siva1 significantly inhibited the invasion and migration of HCT116 cells. It may thus find clinical application for control of cancers.

Keywords

Siva1;cloning;expression;HCT116 cells;invasion;migration

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