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

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. 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

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.

CLC Number:

LIU Yu-hu¹, ZHONG Dong², LIU Juan¹, ZHANG Zhen-shu², CHEN Cun-long², WU Jin-Bao², XIAO Bing², GUO Wen-ying¹. PCR amplification, cloning and protein expression of interferon-inducible transmembrane protein-1 gene[J]. Journal of Southern Medical University, 2005, 25(10): 1221-1224.

References

[1] 刘宇虎,张振书,钟东,等.人源性大肠癌抗原基因噬菌体表达库的构建及酶切鉴定[J].第一军医大学学报,2003,23(6):538-41.Liu YH, Zhang ZS, Zhong D, et al. Construction and identification of the cDNA phage expression library for human colorectal cancer antigens [J]. J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao,2003, 23(6): 538-41.
[2] 刘宇虎,张振书,肖冰,等.人源性大肠癌cDNA噬菌体表达文库的构建及鉴定[J].中华消化杂志,2003,23(10):599-602.Liu YH, Zhang ZS, Xiao B, et al. Construction and identification of human colorectal carcinoma cDNA phage expression library [J].Chin J Dig, 2003, 23(10): 599-602.
[3] 刘宇虎,张振书,钟东,等.人源性大肠癌抗原基因的SEREX筛选[J].世界华人消化杂志,2003,11(9):1378-81.Liu YH, Zhang ZS, Zhong D, et al. SEREX screening of human colorectal carcinoma antigens [J]. World Chin J Digestol, 2003, 11(9): 1378-81.
[4] Hisamatsu T, Watanabe M, Ogata H, et al. Interferon-inducible gene family 1-8U expression in colitis-associated colon cancer and severely inflamed mucosa in ulcerative colitis [J]. Cancer Res,1999, 59(23): 5927-31.
[5] Devito WJ, Stone S, Mori K, et al. Ethanol inhibits prolactin and tumor necrosis factor-alpha, but not gamma interferon-induced expression of intercellular adhesion molecule-1 in human astrocytoma cells [J]. J Cell Biochem, 2000, 77(3): 455-64.
[6] Park SY, Billiar TR, Seol DW. IFN-gamma inhibition of TRAIL-induced IAP-2 upregulation, a possible mechanism of IFN-gammaenhanced TRAIL-induced apoptosis [J]. Biochem Biophys Res Commun, 2002, 291 (2): 233-6.
[7] Lewin AR, Reid LE, McMahon M, et al. Molecular analysis of a human interferon-inducible gene family [J]. Eur J Biochem, 1991,199(2): 417-23.
[8] Takeda K, Okumura K. Current advances and expectations in tumor immunology[J]. Hum Cell, 2001, 14(3): 159-63.

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

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