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采用SMART RACE方法,从东方山羊豆盐诱导抑制性差减杂交cDNA文库中分离到了一个脱水蛋白(GoDHN)基因。该基因cDNA全长1 169bp,开放阅读框843bp,编码281个氨基酸,编码的蛋白质分子量为28.71kDa。经实时荧光定量PCR分析,GoDHN基因在东方山羊豆的茎和叶中表达量明显高于根中表达量,并且基因表达受ABA、NaCl和PEG的诱导,随着诱导时间的增加,表达量呈持续增长趋势。这些结果表明,DHN基因在东方山羊豆的抗逆性中可能起到重要的调控作用。本研究成功构建了pCAMBIA1302-DHN植物表达载体,为下一步转基因研究奠定了基础。
Using the SMART RACE method, a dehydratin (GoDHN) gene was isolated from the cDNA library of Oriental goat bean salt-induced suppression subtractive hybridization. The full-length cDNA of this gene was 1 169 bp in length and 843 bp in open reading frame, encoding a protein of 281 amino acids with a molecular weight of 28.71 kDa. The results of real-time fluorescent quantitative PCR showed that the expression level of GoDHN gene in stem and leaf of oriental goat was significantly higher than that in root, and the expression of GoDHN gene was induced by ABA, NaCl and PEG. With the increase of induction time, Continued growth trend. These results indicate that DHN gene may play an important regulatory role in the resistance of oriental goat. The pCAMBIA1302-DHN plant expression vector was successfully constructed in this study, which laid the foundation for the next transgenic study.