| 茶树黄酮醇合成酶基因的克隆与原核表达 |
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| 引用本文： | 马春雷,陈亮.茶树黄酮醇合成酶基因的克隆与原核表达[J].广西农业生物科学,2009(3):433-438. |
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| 作者姓名： | 马春雷 陈亮 |
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| 作者单位： | 中国农业科学院茶叶研究所茶树资源与改良研究中心;国家茶树改良中心; |
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| 基金项目： | 国家863计划(2006AA10Z171);;现代农业产业技术体系建设专项资金共同资助 |
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| 摘 要： | 本研究采用EST测序技术和RT—PCR技术，获得了一个茶树茶多酚代谢中的重要基因——黄酮醇合成酶（FLS）基因，在GenBank登录（GenBank accession No．EF205150），其序列全长1317bp，其中开放阅读框长996bp，编码331个氨基酸，3]端有一个明显的多聚腺苷酸加尾信号，推测的蛋白分子量约为37．5kD，理论等电点为5．80。序列分析表明它与葡萄FLS基因序列的亲缘关系比较近。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，结果表明茶树黄酮醇合成酶基因能在大肠杆菌BL21中表达，电泳检测到一条大约61kD的外源蛋白，与预测的融合蛋白分子量相符。用Ni-NTA亲和层析柱对融合蛋白进行纯化，得到了纯度在90%以上的纯化蛋白，为进一步研究PET-FLS融合蛋白的活性及功能奠定了基础。
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| 关 键 词： | 茶树 黄酮醇合成酶 基因克隆 序列分析 原核表达 |
Cloning and Prokaryotic Expression of Flavonol Synthase Gene from Tea Plant |
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| Ma Chunlei Chen Liang Research Center for Tea Germplasm , Improvement,Tea Research Institute Chinese Academy of Agricultural Sciences,National Center for Tea Im-provement,Hangzhou.Cloning and Prokaryotic Expression of Flavonol Synthase Gene from Tea Plant[J].Journal of Guangxi Agricultural and Biological Science,2009(3):433-438. |
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| Authors: | Ma Chunlei Chen Liang Research Center for Tea Germplasm Improvement Tea Research Institute Chinese Academy of Agricultural Sciences National Center for Tea Im-provement Hangzhou |
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| Institution: | Ma Chunlei Chen Liang Research Center for Tea Germplasm , Improvement,Tea Research Institute Chinese Academy of Agricultural Sciences,National Center for Tea Im-provement,Hangzhou,310008 |
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| Abstract: | The flavonol synthase gene, which was an important functional gene of catechins biosynthesis pathway, was cloned from tea plant by using EST sequencing and RT-PCR approaches.The full-length cDNA of flavonol synthase gene is 1 317 bp(GenBank accession No.EF205150), containing a 996 bp open reading frame(ORF) encoding a 331 amino acid protein, and its 3' untranslated region has an obvious polyadenylation signal.The deduced protein molecular weight was 37.5 kD and its theoretical isoelectric point was 5.80.Seq... |
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| Keywords: | Tea plant （Camellia sinensis） Flavonol synthase Gene cloning Sequence analysis Prokaryotic expression |
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