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一种新的人细胞色素P450 2A6 cDNA克隆 |
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诸葛坚 钱羽力 谢海洋 余应年
［摘 要］ 目的：克隆人细胞色素P450 2A6 cDNA。方法:用逆转录-PCR和DNA 重组技术, 从人肝组织中扩增人细胞色素P450 2A6基因的cDNA， 并将其连接到pBluescript载体上，对CYP2A6 cDNA进行全序列测定。结果:所克隆的cDNA与Yamano等发表的CYP2A6 cDNA序列相比，在编码区有两个变异，即codon 8 CTG→TTG,编码的氨基酸不变, 为亮氨酸。codon479 GGC(甘氨酸)→GTC(缬氨酸），其余均相同。而在其3′端非编码区变异很多。与Fernandez-Salguero等报道的CYP2A7序列相比，其5′端虽与所克隆的CYP2A6有较多差异，但其codon479也是GTC(缬氨酸）， 在3′端非编码区仅略有差异。而所克隆的CYP2A6 cDNA与Yamano等报道的CYP2A7序列相比，3′端非编码区至所设PCR引物止，所克隆的基因序列完全相同，而在编码区却差异较多。 结论:本实验室克隆的CYP2A6 cDNA 是一种新的CYP2A6 cDNA序列, 有可能来自于新的CYP2A6等位基因的转录。
［主题词］ 细胞色素类；DNA；克隆，分子
［中图分类号］ Q785 ［文献标识码］ A
［文章编号］ 1000-4718(2000)10-0875-04
Cloning of a new human cytochrome P450 2A6 cDNA
ZHU Ge-jian QIAN Yu-li XIE Hai-yang YU Ying-nian
（Department of Pathophysiology and Laboratory of Medical Molecular Biology,
Zhejiang University School of Medicine, Hangzhou 310031,China）
［Abstract］ AIM：To clon human cytochrome P450 2A6 cDNA. METHODS:Using reverse transcription polymerase chain reaction(RT-PCR) and DNA recombinant technique, a full-length cDNA encoding cytochrome P450 2A6(CYP2A6) from human liver was cloned into pBluescript vector. The cDNA segment was identified by DNA sequencing.RESULTS: Comparing with the CYP2A6 sequence, the cloned CYP2A6 cDNA had two different bases, codon 8 CTG(Leu)→TTG(Leu), codon 479 GGC(Gly)→GTC(Val) and was quite different in their 3′end noncoding region. Comparing with CYP2A7 seqence reported by Fernandez-Salguero，the cloned CYP2A6 cDNA had some different in 5′ end coding sequence and several differencey in the 3′ end coding and noncoding sequence, but both codon 479 were GTC(Val).Comparing with the CYP2A7 seqence reported by Yamano，the cloned CYP2A6 cDNA had some difference in the coding sequence but the 3′ end no-coding area was the same. CONCLUSION: The cloned cDNA was a new cDNA of CYP2A6 which may be transcripted from a new al[1] [2] 下一页
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