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AIM:To clone human 15ku selenoprotein geneMETHODS:H9 human T cells were cultured in RPMI1640medium supplemented with 100 mL/L fetal calf serum,mRNAwas isolated from the cells,cDNA library was constructed byRT-PCR.The human 15ku selenoprotein gene was obtainedby PCR and cloned into T vector and sequenced.RESULTS:A unique cDNA fragment about 1 244 bp wasobtained.Sequence analysis identified an open reading framewithin the cDNA.The gene had an in-frame TGA,whichencoded selenocysteine (Sec),and a 3’-UTR SECIS element,which was required for synthesis of selenoprotein.Thepredicted protein molecular mass was about 15ku (162residues).The result was identical with human liver 15kuselenoprotein gene published in Genbank.CONCLUSION: Human 15ku selenoprotein gene can be successfully obtained from T cell line.
AIM: To clone human 15 ku selenoprotein gene METHODS: H9 human T cells were cultured in RPMI 1640 medium supplemented with 100 mL / L fetal calf serum, mRNA was isolated from the cells, cDNA library was constructed by RT-PCR.The human 15 ku selenoprotein gene was obtained by PCR and cloned into T vector and sequenced .RESULTS: A unique cDNA fragment about 1 244 bp wasobtained. Sequence analysis identified an open reading framewithin the cDNA. The gene had an in-frame TGA, whichencoded selenocysteine (Sec), and a 3’-UTR SECIS element, which was required for synthesis of selenoprotein. The predicted protein molecular mass was about 15 ku (162 residues). The result was identical with human liver 15 kuselenoprotein gene published in Genbank. CONCLUSION: Human 15 ku selenoprotein gene can be successfully obtained from T cell line.