光皮桦ACC 氧化酶基因BlACO 的克隆和表达分析

周厚君, 黄华宏, 林二培, 程龙军, 彭沙沙, 童再康*
浙江农林大学林业与生物技术学院, 浙江临安311300

通信作者:童再康;E-mail: zktong@zjfc.edu.cn;Tel: 0571-63732746

摘 要:

ACC (1-aminocyclopropane-1-carboxylic acid)氧化酶(ACO)是植物乙烯合成过程中的关键限速酶, 对乙烯的合成具有 重要的调控作用。以光皮桦茎叶组织提取的 RNA 为模板, 据已报道的 ACO 同源序列设计简并引物, 通过 RT-PCR 扩增获 得部分基因片段, 结合 5', 3' RACE 方法从光皮桦中扩增出 1 个 ACO 的全长 cDNA 序列。该基因 cDNA 全长 1 262 bp, 具有 一个 957 bp 的完整开放阅读框架, 编码含 318 个氨基酸的蛋白。与其他植物中的 ACO 基因进行同源性比对的结果显示, BlACO 蛋白与欧洲白桦的同源性最高, 达到97%。该基因在光皮桦的雄花和雌花中表达量较高, 而在茎中的表达量较低。

关键词:光皮桦; ACO; 基因克隆; 表达

收稿:2010-04-12   修定:2010-07-07

资助:浙江省重大科技专项(2008C02004-1)和浙江省林业科技重点项目(2006SY01)

Cloning and Expression Analysis of ACC Oxidase Gene BlACO in Betula luminifera H. Winkl.

ZHOU Hou-Jun, HUANG Hua-Hong, LIN Er-Pei, CHENG Long-Jun, PENG Sha-Sha, TONG Zai-Kang*
School of Forestry and Biotechnology, Zhejiang Agricultural & Forestry College, Lin’an, Zhejiang 311300, China

Corresponding author: TONG Zai-Kang; E-mail: zktong@zjfc.edu.cn; Tel: 0571-63732746

Abstract:

1-aminocyclopropane-1-carboxylic acid oxidase (ACO) is an important rate-limiting enzyme and plays a critical role in regulation of the ethylene biosynthesis. The BlACO gene was cloned by RT-PCR and 5', 3' RACE methods, using mRNA template from stem and leaf of Betula luminifera, and the designed primers according to the homologous sequences of ACO from other species. Sequence analysis showed that BlACO cDNA was 1 262 bp length and contained a single open reading frame encoding a protein of 318 amino acids. The results of homology comparison showed that BlACO protein shared a 97% homology with Betula pendula. The expression of BlACO gene was higher in male flower and female flower of Betula luminifera, and lower in stem.

Key words: Betula luminifera; ACO; gene clone; expression

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