Journal of Southern Medical University ›› 2005, Vol. 25 ›› Issue (10): 1221-1224.

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PCR amplification, cloning and protein expression of interferon-inducible transmembrane protein-1 gene

LIU Yu-hu1, ZHONG Dong2, LIU Juan1, ZHANG Zhen-shu2, CHEN Cun-long2, WU Jin-Bao2, XIAO Bing2, GUO Wen-ying1   

  1. 1. 东莞市人民医院消化科, 广东, 东莞, 523018;
    2. 南方医科大学南方医院消化病研究所, 广东, 广州, 510515
  • Online:2005-10-20 Published:2005-10-20

Abstract: Objective To study the PCR amplification, cloning and protein expression of interferon-inducible transmembrane protein-1 (IFITMP-1) gene. Methods With the cDNA fragment containing IFITMP-1 gene as template, IFITMP-1 gene was amplified using Pfu enzyme by means of PCR. After EcoRⅠand HindⅢ digestion, the target gene fragment was linked to pUCm-T plasmid and sequenced. The IFITMP-1 gene was cloned into pET-Trx protein expression plasmid, and the condition for protein expression was optimized. Results The length of the PCR product of IFITMP-1 gene-containing cDNA fragment was about 1000 bp. The IFITMP-1 gene was successfully inserted into pUCm-T plasmid with correct sequence and cloning of the IFITMP-1 gene into the pET-Trx protein expression plasmid was achieved. Expression of the fusion protein of pUCm-T plasmid and IFITMP-1 gene was detected after IPTG induction. Conclusion Successful amplification and cloning of the IFITMP-1 gene and its protein expression may facilitate further study of the role of IFITMP-1 gene in colorectal cancer.

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