《植物生理学报》 2016, 52(5): 635-644

中间锦鸡儿CiMYB177的克隆及表达分析

韩晓敏, 邢丹丹, 杨杞, 李国婧, 王瑞刚^*

内蒙古农业大学生命科学学院, 呼和浩特010018

通信作者：王瑞刚；E-mail: wangruigang@imau.edu.cn

摘　要：

MYB转录因子超家族很多基因参与调控生物钟。本文利用RACE技术克隆到一个中间锦鸡儿MYB家族基因CiMYB177, 其cDNA全长2 028 bp, 可以编码433个氨基酸。序列分析发现CiMYB177属于CCA1 (CIRCADIAN AND CLOCK ASSOCIATED1)亚家族, 与大豆的GmMYB177一致性最高(70.3%)。用染色体步移技术克隆到该基因起始密码子上游1 127 bp的启动子序列。用实时荧光定量PCR技术对CiMYB177的转录水平进行分析发现其表达呈节律性的昼夜变化, 并且受干旱、NaCl和冷的诱导。亚细胞定位观察发现CiMYB177定位到细胞核。

关键词：中间锦鸡儿; 生物钟; 基因克隆; MYB转录因子

收稿：2015-11-17   修定：2016-03-28

资助：国家自然科学基金(31360056和31360169)和内蒙古自治区科技创新团队建设计划项目(2015)。 致谢 中国林业科学院齐力旺研究员惠赠了中间锦鸡儿转录组数据。

Cloning and expression analysis of CiMYB177 from Caragana intermedia

HAN Xiao-Min, XING Dan-Dan,YANG Qi, LI Guo-Jing, WANG Rui-Gang^*

College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China

Corresponding author: WANG Rui-Gang; E-mail: wangruigang@imau.edu.cn

Abstract:

Many MYB transcription factors play a role in the circadian clock. In this study, a MYB-related gene CiMYB177 from Caragana intermedia was cloned by RACE, the cDNA sequence was 2 028 bp which encoded 433 amino acids. Sequence analysis showed that CiMYB177 was classified to the CCA1 (CIRCADIAN AND CLOCK ASSOCIATED1) subfamily, and had the highest identity (about 70.3%) with GmMYB177 from Glycine max. Genome walking was used to obtain the 1 127 bp CiMYB177 promoter sequence. The transcriptional expression of CiMYB177 changed diurnally and induced by drought, NaCl and cold. Subcelluar localization revealed that CiMYB177 localized in the nuclei.

Key words: Caragana intermedia; circadian clock; gene cloning; MYB transcription factor