论文部分内容阅读
利用RT-PCR方法从聚乙二醇(PEG)处理的大豆根部组织中克隆了1个大豆乙烯响应因子(ERF)基因,由于其位于大豆基因组第7染色体上,故命名为GmERF7,基因座为Glyma07g02930。序列分析结果表明:该基因含有1个237 bp长的内含子,开放阅读框(ORF)长585 bp,编码194个氨基酸,蛋白质分子量为2.199×104,理论等电点为7.06;通过氨基酸序列比对发现,该序列与其他物种ERF类蛋白质氨基酸序列有很高的相似性;聚类分析结果表明,GmERF7与苜蓿和蓖麻遗传距离最近,与拟南芥和葡萄遗传距离相对较远;半定量RT-PCR分析结果表明该基因主要在大豆叶和豆荚中表达,而且在外源20%PEG处理不同时间后,该基因的表达在根部组织和叶片组织都受到不同程度的诱导,暗示该基因可能对提高植物耐旱性具有一定作用;利用洋葱表皮细胞的亚细胞定位分析结果表明,该蛋白质位于细胞核内,当去除N端信号肽之后,该蛋白质分布于细胞膜部位,这表明该蛋白质在植物体内通过充当转录因子的作用来调节下游基因的表达;适时定量-PCR(Real-time PCR)分析结果证实,该基因在外源ABA处理后呈现先下调后上调又下降的趋势,表明该基因受ABA的诱导,可能参与了ABA依赖的植物抗逆信号转导途径。
One soybean ethylene response factor (ERF) gene was cloned by RT-PCR from polyethylene glycol (PEG) -treated soybean roots. Because of its location on the 7th chromosome of soybean genome, it was named GmERF7. The locus was Glyma07g02930. Sequence analysis showed that the gene contained a 237 bp intron with an open reading frame (ORF) of 585 bp encoding 194 amino acids with a molecular weight of 2.199 × 104 and a theoretical isoelectric point of 7.06. The amino acid sequence The results showed that there was a high similarity between the sequence and the amino acid sequences of ERF proteins in other species. The cluster analysis showed that the genetic distance between GmERF7 and alfalfa and castor was the closest and the genetic distance between Arabidopsis and Arabidopsis was relatively far. The results of quantitative RT-PCR analysis showed that the gene was mainly expressed in soybean leaves and pods. Moreover, the expression of the gene was induced to some extent in roots and leaves at different time points after exogenous 20% PEG treatment, suggesting that the gene may The results of subcellular localization analysis of onion epidermal cells showed that the protein was located in the nucleus. After the N-terminal signal peptide was removed, the protein distributed in the cell membrane, which indicated that the protein was located in the plant Regulated the downstream gene expression by acting as a transcription factor; Real-time PCR analysis confirmed that the gene was treated with exogenous ABA Showing the trend of down-regulation and then up-down, indicating that this gene is induced by ABA and may be involved in ABA-dependent plant stress-resistant signal transduction pathway.