Nan KJ, Li CL, Wei YC, Sui CG, Jing Z, Qin HX, Zhao LJ, Pan BR. Cloning of human 15ku selenoprotein gene from H9 T cells. World J Gastroenterol 2003; 9(8): 1777-1780 [PMID: 12918119 DOI: 10.3748/wjg.v9.i8.1777]
Corresponding Author of This Article
Dr. Chun-Li Li, Department of Medical Oncology, First Hospital of Xi’an Jiaotong University, 1 Jiangkang Road, Xi’an 710061, Shaanxi Province, China. chun128@sohu.com
Article-Type of This Article
Basic Research
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Ke-Jun Nan, Chun-Li Li, Yong-Chang Wei, Chen-Guang Sui, Zhao Jing, Hai-Xia Qin, Department of Medical Oncology, First Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
Li-Jun Zhao, The University of Georgia, Athens, Ga 30602, USA
Bo-Rong Pan, Department of Oncology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China
Author contributions: All authors contributed equally to the work.
Supported by Science and Technology Research and Development Project of Shaanxi Province, No. 2002K10-G1
Correspondence to: Dr. Chun-Li Li, Department of Medical Oncology, First Hospital of Xi’an Jiaotong University, 1 Jiangkang Road, Xi’an 710061, Shaanxi Province, China. chun128@sohu.com
Telephone: +86-29-5324036 Fax: +86-29-5324086
Received: March 3, 2003 Revised: March 10, 2003 Accepted: March 29, 2003 Published online: August 15, 2003
Abstract
AIM: To clone human 15ku selenoprotein gene.
METHODS: H9 human T cells were cultured in RPMI1640 medium supplemented with 100 mL/L fetal calf serum. mRNA was isolated from the cells. cDNA library was constructed by RT-PCR. The human 15ku selenoprotein gene was obtained by PCR and cloned into T vector and sequenced.
RESULTS: A unique cDNA fragment about 1244 bp was obtained. Sequence analysis identified an open reading frame within the cDNA. The gene had an in-frame TGA, which encoded selenocysteine (Sec), and a 3’-UTR SECIS element, which was required for synthesis of selenoprotein. The predicted protein molecular mass was about 15ku (162 residues). The result was identical with human liver 15ku selenoprotein gene published in Genbank.
CONCLUSION: Human 15ku selenoprotein gene can be successfully obtained from T cell line.