《植物生理学报》 2010, 46(1): 5-10

光皮桦中4- 香豆酸辅酶A 连接酶基因Bl4CL 的克隆和表达分析

程龙军*, 童再康, 黄华宏, 楼雄珍

浙江林学院林业与生物技术学院, 浙江临安 311300

通信作者：程龙军；E-mail: ljcheng@zju.edu.cn；Tel: 0571-63742885

摘　要：

以从光皮桦茎叶组织提取的 mRNA 为模板, 根据其他已克隆到的阔叶类树种中 4- 香豆酸辅酶 A 连接酶(4CL)基因的 同源序列设计兼并引物, 进行 RT-PCR 扩增, 获得部分基因片段, 然后结合 5’, 3’ RACE 方法从光皮桦中扩增出 1 个 4CL 基 因的全长 cDNA 序列, 命名为 Bl4CL。该基因 cDNA 全长为 1 983 bp (GenBank 登录号 FJ410448), 具有完整的开放阅读框架 (69~1 697 bp), 编码蛋白为 542 个氨基酸, 包含一个 AMP 结合功能域和一个含有 12 个氨基酸的功能基序。与其他植物中 的4CL 进行同源性比对的结果显示, Bl4CL 蛋白与东北白桦的同源性最高, 达到了98%。该基因在光皮桦的根和茎中表达 量较高, 而在花和叶中的表达量低。

关键词：光皮桦; 4- 香豆酸辅酶 A 连接酶; 基因克隆; 表达

收稿：2009-09-21   修定：2009-12-22

资助：浙江省重大科技专项(2008C02004-1)

Cloning and Expression Analysis of 4-coumarate: CoA Ligase Gene Bl4CL in Betula luminifera H. Winkl.

CHENG Long-Jun*, TONG Zai-Kang, HUANG Hua-Hong, LOU Xiong-Zhen

School of Forestry and Biotechnology, Zhejiang Forestry College, Lin’an, Zhejiang 311300, China

Corresponding author: CHENG Long-Jun; E-mail: ljcheng@zju.edu.cn; Tel: 0571-63742885

Abstract:

A 4-coumarate: CoA ligase gene (Bl4CL) was amplified by RT-PCR and 5’, 3’ RACE methods, when mRNA from stem and leaf of Betula luminifera were used as a template, and the primers were designed according to the homologous sequences of 4CL from broadleaf species. Sequence analysis showed that Bl4CL cDNA was 1 983 bp and contained a single open reading frame (69–1 697 bp) encoding a protein of 542 amino acids (GenBank No. FJ410448). The protein had a motif of 12 amino acids and a functional AMP-binding domain. The results of homology comparison showed that Bl4CL protein shared a 98% homology with Betula platyphylla. And the expression of Bl4CL gene were high in roots and stems of Betula luminifera, and low in leaves and flowers.

Key words: Betula luminifera; 4-coumarate: CoA ligase; gene clone; expression