Carcinogenesis, Teratogenesis & Mutagenesis ›› 2010, Vol. 22 ›› Issue (3): 182-185.doi: 10.3969/j.issn.1004-616x.2010.03.006

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Construction of p53 polymorphic plasmid by site-directed mutagenesis

QIU Shi1;2;CAI Yun1;GAO Xing1;GU Shou-zhi3;LIU Ze-jun1   

  1. 1. Southwest Cancer Center, Southwest Hospital Third Military Medical University, Chongqing 400038； 2. Biotherapy Center, General Hospital of Beijing Military Area Commond of China PLA, Beijing 100068, China; 3. School of Rehabilitation Sciences, Seirei Christopher University, Hamamatsu 433－8558, Japan
  • Received:2009-10-30 Revised:2010-03-10 Online:2010-05-30 Published:2010-05-30
  • Contact: Liu Ze-jun

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Abstract: OBJECTIVE: To investigate the function of p53 polymorphic variants, we constructed the p53 codon 72 polymorphic plasmid. METHODS: Primers carrying the appropriate mutation were employed in a two-step PCR amplification. The mutated PCR products were subsequently cloned into pReceiver-M01 expression vector. The p53 protein translated in vitro was used to interact with iASPP to certify its biological activity. RESULTS: The sequence of the recombinant eukaryotic expression vector containing CCC to CGC mutation was proved by DNA sequencing. This p53 polymorphic variant could interact with iASPP, implying its biological activitical. CONCLUSION: The recombinant p53 eukaryotic expression vector containing codon 72 polymorphism was constructed successfully, laying a foundation for further studies on the function of p53.

Key words: p53 polymorphism, site-directed mutagenesis, overlap extension PCR

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