茶树黄酮合成酶Ⅱ基因全长cDNA序列的克隆和实时荧光定量PCR检测

doi:10.13305/j.cnki.jts.2009.5.004

茶叶科学 ›› 2009, Vol. 29 ›› Issue (5): 347-354.doi: 10.13305/j.cnki.jts.2009.5.004

茶树黄酮合成酶Ⅱ基因全长cDNA序列的克隆和实时荧光定量PCR检测

乔小燕^1,2, 马春雷¹, 陈亮^1,*

1. 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,浙江杭州 310008;
2. 广东省农业科学院茶叶研究所,广东广州,510640

收稿日期:2009-02-12 修回日期:2009-04-09 出版日期:2009-10-15 发布日期:2019-09-09
通讯作者: * liangchen@mail.tricaas.com
作者简介:乔小燕（1982— ）,女,山西繁峙人,主要从事茶树分子生物学研究。
基金资助:
国家863计划（2006AA10Z171）和“现代农业产业技术体系建设专项资金资助”内容之一

Molecular Cloning and Real-Time PCR Analysis of Flavone SynthaseⅡ Gene Full-length cDNA from the Tea Plant

QIAO Xiao-yan^1,2, MA Chun-lei¹, CHEN Liang^1,*

1. Research Center for Tea Germplasm and Improvement, Tea Research Institute of the Chinese Academy of Agricultural Sciences; National Center for Tea Improvement, Hangzhou 310008, China;
2. Tea Research Institute of the Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

Received:2009-02-12 Revised:2009-04-09 Online:2009-10-15 Published:2019-09-09

摘要/Abstract

摘要： 利用RT-PCR和RACE技术,克隆得到茶叶中合成黄酮类化合物的关键酶基因—黄酮合成酶Ⅱ,该基因属于细胞色素P450家族,在GenBank登录号为FJ169499。黄酮合成酶ⅡcDNA全长序列1 824 bp,其中1 605 bp为编码区,编码534个氨基酸,分子量为60.4 kD。通过SYBR Green I实时荧光定量PCR检测,发现黄酮合成酶Ⅱ在茶树一芽二叶春梢、当季成熟叶、花瓣、花蕊、种子中都有表达,成熟叶的表达量显著高于其他四种组织。

关键词: 茶树, 黄酮合成酶Ⅱ, 实时荧光PCR, 细胞色素P450家族

Abstract: Using RT-PCR and RACE techniques, flavone synthase Ⅱ (FSⅡ) gene, the key enzyme in synthesize flavone, was cloned in the tea plant. The FS II belongs to cytochrome P450 superfamily. The GenBank Accession No. is FJ169499 in the NCBI. The FSⅡ gene cDNA had 1 824 bp in full-length with 1 605 bp in coding region, encoding 534 amino acids. The putative molecular weight was 60.4 kD. The expression levels of flavone synthaseⅡ gene in spring ‘two and a bud’, mature leaves, petals, stamen and pistils, and seeds of the tea plant were assessed using SYBR Green I Real-time PCR. The expression in mature leaves was significantly higher than that in the other four tissues.

Key words: tea plant, flavone synthaseⅡ, Real-Time PCR, cytochrome P450 superfamily

中图分类号:

S571.1
Q523

乔小燕, 马春雷, 陈亮. 茶树黄酮合成酶Ⅱ基因全长cDNA序列的克隆和实时荧光定量PCR检测[J]. 茶叶科学, 2009, 29(5): 347-354. doi: 10.13305/j.cnki.jts.2009.5.004.

QIAO Xiao-yan, MA Chun-lei, CHEN Liang. Molecular Cloning and Real-Time PCR Analysis of Flavone SynthaseⅡ Gene Full-length cDNA from the Tea Plant[J]. Journal of Tea Science, 2009, 29(5): 347-354. doi: 10.13305/j.cnki.jts.2009.5.004.

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茶树黄酮合成酶Ⅱ基因全长cDNA序列的克隆和实时荧光定量PCR检测

Molecular Cloning and Real-Time PCR Analysis of Flavone SynthaseⅡ Gene Full-length cDNA from the Tea Plant

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