《植物生理学报》 2009, 45(8): 765-770

柠条锦鸡儿肌动蛋白基因的克隆和表达稳定性分析

王学敏, 王美珍, 王赞, 高洪文

中国农业科学院北京畜牧兽医研究所, 北京100193

通信作者：高洪文；E-mail: gaohongwen@263.net；Tel: 010-62894560

摘　要：

以几种豆科植物中肌动蛋白的氨基酸保守序列设计简并引物, 采用 RT-PCR 结合 RACE 扩增技术, 从柠条锦鸡儿叶片中克隆到一个编码肌动蛋白的基因, 命名为 CkACT。该基因 cDNA 全长为 1655 bp, 开放阅读框 1134 bp, 编码 377 个氨 基酸。氨基酸比对分析表明, 该基因编码的氨基酸与其他植物肌动蛋白基因具有较高的同源性。不同组织中的表达基本上 一致, 不同发育时期的基因表达相似, 低温、NaCl、干旱和 ABA 处理后的表达强度没有明显差异, 说明 CkACT 基因表达 稳定。

关键词：肌动蛋白; 柠条锦鸡儿; 克隆; 表达稳定性

收稿：2009-03-09   修定：2009-05-22

资助：国家“ 十一五” 科技支撑计划(2008BADB3B01,2006BAD01A16 )和中国农业科学院基本科研业务费(Ywf-td-3)。

Clone and Expression Stability Analysis of Actin Gene in Caragana korshinskii

WANG Xue-Min, WANG Mei-Zhen, WANG Zan, GAO Hong-Wen*

Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100193, China

Corresponding author: GAO Hong-Wen; E-mail: gaohongwen@263.net; Tel: 010-62894560

Abstract:

An actin gene, CkACT, was cloned from the leaves of Caragana korshinskii by using RT-PCR and RACE with degenerate primers which were synthesized based on the high homologous region among the sequences of several leguminous ACT genes. The full-length cDNA of CkACT was 1 655 bp and contained a 1 134-bp open reading frame encoding a 377-amino acid protein. The deduced amino acid sequence of CkACT protein shared high identity with other ACTINs via multiple alignment. CkACT expressed constitutively in all tested organs, including root, shoot and leaf. There was no significant difference in mRNA expression in different development stage or under cold, NaCl, drought and ABA treatment. These results indicated that the expression of CkACT is stable.

Key words: actin; Caragana korshinskii; cloning; expression stability