关键词: p53基因, 基因克隆, 原核表达

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