J4 ›› 2009, Vol. 47 ›› Issue (05): 1077-1080.

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Cloning and Prokaryotic Expression of p53 Gene

WANG Ji, GAO Ruijuan, YUAN Dawei, WANG Li   

  1. Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China
  • Received:2008-12-22 Online:2009-09-26 Published:2009-11-03
  • Contact: WANG Li E-mail:wangli@nenu.edu.cn.

Abstract:

p53 cDNA was obtained from human peripheral blood lymphocytes by RTPCR, and then the p53 fragments were inserted into pQE40 vector to construct the recombinant plasmid pQE40p53. Furthermore, pQE40p53 plasmids were transformed into E.coli M15, and then induced by 1 mmol/L IPTG to express p53 protein in E.coli M15 expression system. The recombinant p53 protein fused with 6His-tag was purified by Ni-NAT resin affinity chromatography, and then urea was removed with
gradual dialysis. Western Blot confirmed that purified 6His-p53 recombinant protein was  obtained.

Key words: p53 gene, gene cloning, prokaryotic expression

CLC Number: 

  • Q78