紫花苜蓿GDP解离抑制蛋白基因cDNA的克隆、分析及烟草转化

采用RT-PCR方法,克隆得到紫花苜蓿(Medicago sativa L.)GDP解离抑制蛋白基因cDNA核心区域序列,命名为MsGDI1。序列分析结果表明,该cDNA核心区域全长1575 bp,包含一个1332 bp的最大开放阅读框,编码444个氨基酸。半定量RT-PCR分析结果表明,在盐胁迫和铝胁迫条件下,MsGDI1基因的表达随着胁迫时间的增加而上升,该基因可能与紫花苜蓿抗盐耐铝机理有关。借助于新构建的该基因超表达载体,用农杆菌介导方法转化烟草(Nicotiana tabacum L.)。转基因烟草植株PCR检测的结果表明,MsGDI1基因已经成功插入到烟草基因组中,为进一步研究该基因的功能奠定了基础。

Cloning and Characterization of a GDP Dissociation Inhibitor Protein Gene from Medicago sativa L.and Expression in Tobacco

A cDNA core region of alfalfa GDP dissociation inhibitor protein Gene named MsGDI1 was isolated by RT-PCR.Sequence analysis reveals that the full length of the core region is 1575 bp and contains 1332 bp open reading frame(444 amino acids).Semi-quantitative RT-PCR analysis show that the expression level of MsGDI1 increases significantly with salt-or Aluminum-stress increasing.This result indicates that MsGDI1 may play an important role in both salt and Aluminum tolerances of Medicago sativa L..The MsGDI1 gene is introduced into tobacco by Agrobacterium mediated transformation system using overexpression vector.PCR analyses show that transgenic tobaccos contain the MsGDI1 gene.The function of MsGDI1 gene will be further studied.