从H9 T细胞中克隆人15ku硒蛋白基因。
Cloning of human 15ku selenoprotein gene from H9 T cells.
作者信息
Nan Ke-Jun, Li Chun-Li, Wei Yong-Chang, Sui Chen-Guang, Jing Zhao, Qin Hai-Xia, Zhao Li-Jun, Pan Bo-Rong
机构信息
Department of Medical Oncology, First Hospital of Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China.
出版信息
World J Gastroenterol. 2003 Aug;9(8):1777-80. doi: 10.3748/wjg.v9.i8.1777.
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 1 244 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.
目的
克隆人15ku硒蛋白基因。
方法
将H9人T细胞培养于补充有100 mL/L胎牛血清的RPMI1640培养基中。从细胞中分离mRNA。通过RT-PCR构建cDNA文库。通过PCR获得人15ku硒蛋白基因，并克隆到T载体中进行测序。
结果
获得了一个约1244 bp的独特cDNA片段。序列分析确定该cDNA内有一个开放阅读框。该基因有一个框内TGA，其编码硒代半胱氨酸（Sec），还有一个3'-UTR SECIS元件，这是硒蛋白合成所必需的。预测的蛋白质分子量约为15ku（162个氨基酸残基）。结果与Genbank中公布的人肝脏15ku硒蛋白基因一致。
结论
可从T细胞系中成功获得人15ku硒蛋白基因。
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