东方山羊豆脱水蛋白基因的克隆及初步分析  

Cloning and analysis of the dehydrin(DHN) gene from Galega orientalis

作　　者：李玉坤[1] 王学敏[1] 王赞[1] 李俊[1] Vladimir C Nikolay D 孙桂枝[1] 高洪文[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]俄罗斯瓦维洛夫全俄植物栽培研究所,圣彼得堡190000

出　　处：《草业学报》2012年第1期176-183,共8页Acta Prataculturae Sinica

基　　金：国际科技合作与交流专项(2010DFR30620-2);现代农业产业技术体系建设专项牧草产业体系资金资助

摘　　要：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 rapid amplification of cDNA ends（RACE） method,a cDNA clone encoding dehydrin（GoDHN） was isolated from a cDNA SSH（suppression subtractive hybridization） library induced by salt stress from Galega orientalis.The full-length of cDNA sequence was 1 169 bp and included an 843 bp open reading frame.Molecular weight of this protein was 28.71 kDa.Results of real-time PCR indicated that the expression of quantity of GoDHN was higher in leaves and stems than in roots.Abiotic stress,such as NaCl and PEG,all induced the expression of GoDHN.Application of the exogenous hormone ABA also up-regulated mRNA accumulation.The expression of GoDHN increased with the induction time.These results indicated that the DHN gene may be involved in regulation of stress-resistance in G.orientalis.Construction of a pCAMBIA1302-DHN vector formed a basis for gene transformation.

关 键 词：东方山羊豆 脱水蛋白基因 基因克隆 实时荧光定量PCR 植物表达载体 

分 类 号：S54[农业科学—作物学]