《植物生理学报》 2009, 45(12): 1160-1166

小黑杨环锌指蛋白基因的克隆与表达分析

王雷, 周博如, 吴丽丽, 吕澈妍, 曲跃军, 郑威, 姜廷波*

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：姜廷波；E-mail: tbjiang@yahoo.com；Tel: 0451-82190607

摘　要：

用 cDNA-AFLP 技术从小黑杨中克隆与盐胁迫反应相关的 cDNA 片段, 进一步应用 RACE 技术克隆出具有完整开放 读码框的小黑杨环锌指蛋白基因(PsnRZF), 该基因全长 1 061 bp, 其中 5' 非翻译区为 184 bp, 3' 非翻译区为 82 bp, 开放读码 框为 795 bp, 编码264 个氨基酸, 预测蛋白的分子量为30.25 kDa, 理论等电点为8.04。实时定量PCR 检测的结果显示, 正常 生长条件下该基因在根、茎、叶中都表达; NaCl 胁迫下, 该基因在根、茎、叶中的表达量升高。在叶中的表达量随着处理 时间的延长而逐渐升高, 胁迫处理后第6天表达量达到最高。

关键词：小黑杨; 环锌指蛋白; RACE; 实时定量PCR

收稿：2009-09-08   修定：2009-11-30

资助：黑龙江省重点科技攻关项目(GB06B303-5)。

Cloning and Expression Analysis of a Ring Zinc-Finger Gene in Populus simonii×P. nigra

WANG Lei, ZHOU Bo-Ru, WU Li-Li, LÜ Che-Yan, QU Yue-Jun, ZHENG Wei, JIANG Ting-Bo*

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

Corresponding author: JIANG Ting-Bo; E-mail: tbjiang@yahoo.com; Tel: 0451-82190607

Abstract:

For studies of differential gene expression, cDNA-AFLP was applied in Populus simonii×P. nigra under salt-stress. The 1 061 bp full length cDNA of ring zinc-finger gene was isolated by rapid amplification of cDNA ends (RACE), including a 184 bp 5' untranslated region, an 82 bp 3' untranslated region and a 795 bp open reading frame encoding 264 amino acid residues. The molecular weight of deduced protein was 30.25 kDa with a theoretical pI of 8.04. Real-time PCR revealed that PsnRZF gene was expressed in roots, stems and leaves under normal growth, and the expression level was increased under salt-stress. The expression level of PsnRZF gene in leaves was increased gradually and the peak value was appeared at the 6th day under NaCl stress.

Key words: Populus simonii×P. nigra; ring zinc finger; RACE; real-time PCR