| 茶树咖啡酸氧甲基转移酶基因的克隆及原核表达 |
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| 引用本文： | 李小霞,余有本.茶树咖啡酸氧甲基转移酶基因的克隆及原核表达[J].西北农业学报,2011,20(5):139-143. |
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| 作者姓名： | 李小霞 余有本 |
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| 作者单位： | 西北农林科技大学园艺学院,陕西杨凌,712100 |
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| 摘 要： | 以从茶树叶子中提取的总RNA为模板,结合RT-PCR利用RACE技术,得到咖啡酸氧甲基转移酶(caffeic acid o-methyltransferase,COMT)基因全长1 381bp,其开放阅读框为1 092bp,编码393个氨基酸,推测的蛋白分子量约为39.661 9ku,理论等电点为5.66,在GenBank的登录号为HM204933。其核苷酸序列与欧洲白桦、苎麻、中果咖啡和毛果杨的COMT基因相似性分别为80%、78%、77%、77%,氨基酸序列与毛果杨的COMT基因达100%。将该基因重组到表达载体pET-32a(+)中进行原核表达,经IPTG诱导,SDS-PAGE检测结果表明茶树COMT基因能在大肠杆菌BL21(DE3)中表达,电泳检测到一条大约60ku的外源蛋白,与预测的融合蛋白分子量相符。
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| 关 键 词： | 茶树 咖啡酸氧甲基转移酶 RACE 原核表达 |
Cloning and Prokaryotic Expression of Genecoding for Caffeic Acid o methyltransferase (COMT) from Tea |
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| LI Xiaoxia and YU Youben.Cloning and Prokaryotic Expression of Genecoding for Caffeic Acid o methyltransferase (COMT) from Tea[J].Acta Agriculturae Boreali-occidentalis Sinica,2011,20(5):139-143. |
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| Authors: | LI Xiaoxia and YU Youben |
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| Institution: | LI Xiaoxia and YU Youben(College of Horticulture,Northwest A&F University,Yangling Shaanxi 712100,China) |
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| Abstract: | A cDNA encoding caffeic acid o-methyltransferase(COMT) was cloned by the methods of RT-PCR and rapid amplification of cDNA ends(RACE) using the total RNA isolated from the leaf of tea as template,which encodes 393 amino acids with a theoretical molecular weight of 39.661 9 ku and an isoelectric point of 5.84.The accession number is HM204933.The cDNA sequence shows a significant homology to the COMT genes from Betula pendula(80%),Boehmeria nivea(78%),Coffea canephora(77%) and Populus trichocarpa(77%).The ami... |
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| Keywords: | Tea Caffeic acid o-methyltransferase(COMT) RACE Prokaryotic expression |
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