| 杨树木质素合成酶hct基因的克隆及核苷酸序列分析 |
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| 引用本文： | 王雪霞,曹方,薛永常.杨树木质素合成酶hct基因的克隆及核苷酸序列分析[J].辽宁林业科技,2009(6):4-7. |
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| 作者姓名： | 王雪霞 曹方 薛永常 |
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| 作者单位： | 1. 大连工业大学,生物与食品工程学院,辽宁省发酵工程重点实验室,辽宁,大连,116034;石药集团中奇制药技术(石家庄)有限公司,河北石家庄052160
2. 大连工业大学,生物与食品工程学院,辽宁省发酵工程重点实验室,辽宁,大连,116034 |
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| 摘 要： | 根据毛果杨全序列（AC185363.2）中唯一具有转移酶功能的保守区设计引物,以正在分化的2年生欧美杨107次生木质部总RNA为模板经RT-PCR扩增出hct基因片段,与pMD20-T载体连接,重组质粒经特异引物扩增、限制性内切酶酶切和测序鉴定。结果表明,扩增片段长度为709bp,包含一个708bp的开放阅读框,与毛果杨全序列及HCT2（EU603314）的同源性均为98%,编码的氨基酸序列与毛果杨HCT2氨基酸（ACC63883.1）的同源性为98%,断定我们克隆的cDNA为hct,属于转移酶超家族,GenBank登录号为FM202091,该重组质粒命名为pMD20-T-hct。
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| 关 键 词： | 木质素 hct基因 cDNA克隆 序列分析 |
Cloning and sequence analysis of lignin biosynthetic enzyme hct gene in Poplar |
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| WANG Xue-xia,CAO Fang,XUE Yong-chang.Cloning and sequence analysis of lignin biosynthetic enzyme hct gene in Poplar[J].Liaoning Forestry Science and Technology,2009(6):4-7. |
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| Authors: | WANG Xue-xia CAO Fang XUE Yong-chang |
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| Institution: | WANG Xue-xia, CAO Fang, XUE Yong-chang(1.The Key Laboratory for Fermentation Engineering in Liaoning Province,College of Biology and Food Engineering,Dalian Polytechnic University,Dalian 116034,China;2.Zhongqi Pharmacy in Shijiazhuang,Pharmaceutical Group Limited,Shijiazhuang 052160,China) |
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| Abstract: | Based on the full sequence conserved domain（transferase）of Populus trichocarpa（accession number：AC185363.2）,a pair of primers was designed and used to amplify a fragment of hct（shikimate/quinate hydroxycinnamoyltransferase）gene by RT-PCR from poplar（Populus×euramericana cv.‘74/76 ）developing second xylem RNA,then cloned into pMD20-T vector,identified by PCR amplification,restriction enzyme cutting, sequencing and named pMD20-T-hct.The sequence of the amplified DNA fragment was 709bp, containing a 708bp ORF （open reading frame）. The identity of the fragment with Populus trichocarpa complete sequence and HCT2 both are 98%. Alignment with amino acid ofPopulus triehocarpa HCT2 showed that their homology was 98%. Therefore, we confirmed that the cloned DNA fragment was hct, belonged to transferase superfamily. It was submitted to GenBank and the accession number was FM202091 and named PMD20-T-hct. |
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| Keywords: | lignin shikimate/quinate hydroxycinnamoyltransferase cDNA cloning sequence analysis |
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