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茶叶科学 ›› 2007, Vol. 27 ›› Issue (2): 127-132.doi: 10.13305/j.cnki.jts.2007.02.006

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茶树查尔酮异构酶基因克隆及序列分析

马春雷1,2,赵丽萍1,张亚丽1,2,陈亮1*   

  1. 1. 中国农业科学院茶叶研究所茶树资源和遗传改良与分子生物学实验室,浙江 杭州 310008;
    2. 中国农业科学院研究生院,北京 100081
  • 出版日期:2007-06-25 发布日期:2019-09-11
  • 通讯作者: *陈亮,liangchen@mail.tricaas.com
  • 作者简介:马春雷(1982— ),男,黑龙江人,硕士研究生,从事茶树分子生物学研究。
  • 基金资助:
    国家863计划(2006AA10Z171)、浙江省“钱江人才”计划项目(2006R10042)、浙江省重点科技项目(2004C22033)、人事部和教育部留学回国人员科研基金(2005-134,2005-383)内容之一

Molecular Cloning and Sequence Analysis of Chalcone Isomerase Gene of Tea Plant (Camellia sinensis)

MA Chun-lei1,2, ZHAO Li-ping1, ZHANG Ya-li1,2, CHEN Liang1,*   

  1. 1. Lab for Tea Germplasm, Genetic Improvement and Molecular Biology, Tea Research Institute Chinese Academy of Agricultural Sciences, Hangzhou 310008, China;
    2. Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Online:2007-06-25 Published:2019-09-11

摘要: 茶叶是世界上最流行的无酒精饮品之一,含有许多有价值的次生代谢产物,如儿茶素类物质。分离和克隆重要的茶树功能基因,对利用基因工程技术进行茶树遗传调控具有重要意义。本文采用EST测序技术和T4RNA连接酶介导的5′RACE技术,获得了一个茶树儿茶素代谢中的重要基因—查尔酮异构酶(CHI)基因,在GenBank登录(DQ904329),其序列全长1 163 bp,其中开放阅读框长723 bp,编码240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26.4 kD,理论等电点为5.19。序列分析表明它与番茄CHI基因序列的亲缘关系比较近。

关键词: 茶树, 查尔酮异构酶, 基因克隆, 序列分析

Abstract: Tea is one of the most popular non-alcoholic healthy beverages in the world, which possesses great value as a source of secondary metabolic products, such as catechins. Isolation and cloning of important functional genes of tea plant (Camellia sinensis) is of crucial significance for using biotechnology method to regulate the metabolism of tea plant. In this paper, the chalcone isomerase gene, which was an important functional gene of catechins biosynthesis pathway, was cloned from tea plant by using EST sequencing and RACE (rapid amplification of cDNA ends) approaches. The full-length cDNA of chalcone isomerase gene is 1 163 bp (GenBank Accession No. DQ904329), containing a 723bp open reading frame (ORF) encoding a 240 amino acid protein, and its 3′ untranslated region has an obvious polyadenylation signal. The deduced protein molecular weight was 26.4 kD and its theoretical isoelectric point was 5.19. Sequence analysis result showed that it is closely related with that of Lycopersicon esculentum.

Key words: tea plant (Camellia sinensis), chalcone isomerase, gene cloning, sequence analysis

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