《植物生理学报》 2009, 45(4): 323-328

二色补血草OEE2 基因的克隆和表达

张大伟, 王玉成*, 杨传平

东北林业大学林木遗传育种与生物技术教育部重点实验室, 哈尔滨150040

通信作者：王玉成；Tel: 2008-11-21

摘　要：

从二色补血草中分离出一条含有完整开放读码框(ORF)序列的 OEE2 基因。该基因全长 994 bp, 其中 5' 非翻译区 27 bp, 3' 非翻译区 160 bp, ORF 全长 807 bp, 共编码 264 个氨基酸, 编码蛋白的分子量为 28.2 kDa, 理论上的等电点为 7.66。 BlastP 分析表明, 二色补血草OEE2 与马铃薯OEE2 序列同源性最高, 与喇叭水仙OEE2 序列同源性最低, 从9 个物种的氨基 酸多序列比对中可以看出, OEE2 的氨基酸序列保守性较高。实时定量RT-PCR 方法检测该基因对低温、NaCl和聚乙二醇 (PEG)胁迫的基因表达模式的结果表明, PEG和低温能诱导OEE2基因在二色补血草叶中表达, 这两种处理的OEE2基因表 达量于胁迫48 h 后都达到高峰, 而在NaCl胁迫下OEE2 在二色补血草根和叶中表达都受抑制。

关键词：二色补血草; OEE2; 基因表达; 实时定量RT-PCR

收稿：2008-11-21   修定：2009-02-23

资助：教育部科学技术研究重点项目(1070 37)、国家自然科学基金面上项目(30571509)和黑龙江省攻关重点项目(GB06B303-1)。

Cloning and Expression Analysis of An Oxygen-evolving Enhancer Protein II Gene (OEE2) from Limonium bicolor (Bunge) Kuntze

ZHANG Da-Wei, WANG Yu-Cheng*, YANG Chuan-Ping

Key Laboratory of Forest Tree Genetic Breeding and Biotechnology, Ministry of Education, Northeast Forestry University, Harbin 150040, China

Corresponding author: WANG Yu-Cheng; Tel: 2008-11-21

Abstract:

The sequence of oxygen-evolving enhancer protein II (OEE2), which contained a complete open reading frame (ORF), was cloned from a cDNA library of Limonium bicolor. The sequence was 994 bp in length, including a 27 bp sequence of 5' untranslated region and a 160 bp sequence of 3' untranslated region. It had an ORF of 807 bp in length, which encoded a deduced amino acids of 264 residues. The molecular weight of deduced protein was 28.2 kDa with a theoretical pI of 7.66. Multiple sequence alignments of OEE2 proteins from 9 plant species revealed that the OEE2 proteins shared high identities in amino acid sequence. The BlastP analysis revealed that amino acid sequence of OEE2 protein from L. bicolor shared the highest amino acid sequence similarity with that from Solanum tuberosum and the lowest similarity with that from Narcissus pseudonarcissus among the 9 OEE2 proteins. We examined the expression pattern of the OEE2 gene in leaves and roots of L. bicolor in response to NaCl, low temperature and PEG stresses at different time points by using real time RT-PCR. The results showed that OEE2 could be induced by low temperature and PEG treatment in leaves, its expression reached maximum level after treatment for 48 h by both low temperature and PEG. However, the expression of OEE2 was inhibited by NaCl treatment in both leaves and roots of L. bicolor.

Key words: Limonium bicolor; oxygen-evolving enhancer protein 2; gene expression; real time RT-PCR