小黑杨环锌指蛋白基因的克隆与表达分析

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

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1061bp,其中5'非翻译区为184bp,3'非翻译区为82bp,开放读码框为795bp,编码264个氨基酸,预测蛋白的分子量为30.25kDa,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。

For studies of differential gene expression, cDNA-AFLP was applied in Populus simoniixP, 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.