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Cloning of human 15ku selenoprotein gene from H9 T cells
Ke-Jun Nan1, Chun-Li Li, Yong-Chang Wei
1Department of Medical Oncology, First Hospital of Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China.
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.
