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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。
The cDNA fragment related to salt stress response was cloned from Populus simonii by cDNA-AFLP technique. The full open reading frame (ORF) gene of PsnRZF was cloned by RACE technique. The full length cDNA of PsnRZF was 1061 bp . The 5 ’untranslated region was 184 bp, the 3’ untranslated region was 82 bp, the open reading frame was 795 bp, encoding 264 amino acids. The predicted molecular weight was 30.25 kDa and the theoretical isoelectric point was 8.04. The results of real-time quantitative PCR showed that the gene was expressed in roots, stems and leaves under normal growth conditions. Under NaCl stress, the gene expression in roots, stems and leaves increased. The expression level in leaves gradually increased with the prolongation of treatment time, reaching the highest level on the 6th day after stress treatment.