| 茶树咖啡因合成酶基因RNA干涉表达载体构建 |
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| 引用本文： | 张广辉,梁月荣,陆建良,董俊杰.茶树咖啡因合成酶基因RNA干涉表达载体构建[J].茶叶科学,2006,26(4):243-248. |
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| 作者姓名： | 张广辉 梁月荣 陆建良 董俊杰 |
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| 作者单位： | 浙江大学茶学系,浙江,杭州,310029;江苏省徐州生物工程学校,江苏,徐州,221006;浙江大学茶学系,浙江,杭州,310029 |
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| 基金项目： | 国家自然科学基金;浙江省科技计划 |
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| 摘 要： | 咖啡因合成酶（TCS）是茶树咖啡因生物合成途径中的一个关键酶,催化7-甲基黄嘌呤转变为可可碱和可可碱转变为咖啡因。抑制TCS基因表达是培育低咖啡因茶树的最有效途径。用RT-PCR方法扩增茶树TCS基因的两个cDNA片段,并克隆入T-载体中。干涉载体和TCS基因片段的T克隆经限制性内切酶两次双酶切与连接,将两个TCS基因片段分别反向重复插入到干涉载体pFGC5941,构建了TCS基因的RNA干涉载体,分别命名为pFGC5941-TCS02和pFGC5941-TCS03。经PCR和DNA测序获得验证。TCS基因RNA干涉载体的构建为培育低咖啡因茶树奠定了基础。
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| 关 键 词： | 茶树 咖啡因合成酶基因 RNA干涉 PCR |
| 文章编号： | 1000-369X（2006）04-243-06 |
| 收稿时间： | 2006-04-11 |
| 修稿时间： | 2006-09-13 |
Construction of Tea Caffeine Synthase Gene RNAi Vector |
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| ZHANG Guang-hui,LIANG Yue-rong,LU Jian-liang,DONG Jun-jie.Construction of Tea Caffeine Synthase Gene RNAi Vector[J].Journal of Tea Science,2006,26(4):243-248. |
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| Authors: | ZHANG Guang-hui LIANG Yue-rong LU Jian-liang DONG Jun-jie |
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| Institution: | 1. Department of Tea Science, Zhejiang University, Hangzhou 310029, China;2. Xuzhou Higher Vocational School of Bioengineering, Xuzhou 221006, China |
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| Abstract: | Tea Caffeine synthase (TCS) is one of the key enzymes involved in caffieine biosynthsis in tea plant (Camellia sinensis),which catalyses conversions of 7-methylxanthine to theobromine and theobromine to caffeine. Inhibition of TCS gene expression can leads to breeding low caffeine tea cultivars. Two cDNA fragments of TCS gene were amplified by RT-PCR, and ligated into T-vector. The two TCS gene fragments were inserted into RNAi vector pFGC5941 in reverse direction after double digestion with two pairs of restriction endonucleases. The insertion of two fragments, namely pFGC5941-TCS02 and pFGC5941-TCS03, into the RNAi vector were confirmed by PCR and DNA sequencing. |
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| Keywords: | PCR |
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