大豆α′基因在烟草植株中的表达

EXPRESSION OF α′-SUBUNIT GENE OF SOYBEAN IN TOBACCO PLANTS

将大豆种子7s 贮藏蛋白α'亚基基因通过含改装 Ti 质粒的根癌农杆菌(Agrobacteriumtumefaciens)感染转化烟草叶片,并使转化组织再生成完整植株。所有再生的转基因烟草植株正常开花结籽,形在态上未发生任何变化。Southern 杂交证明,α'亚基基因已插入烟草基因组中。ELISA 分析表明,转基因植株种子中α'亚基蛋白的含量显著高于非转化植株,而在叶和茎中α'亚基蛋白的含量转化植株和非转化植株并无差异。说明α'亚基基因在转化植株中可以表达,但只在种子中积累,而不在叶片和茎中积累。由此可见,α'亚基基因在转基因烟草中的表达和积累方式与在大豆中的表达和积累方式是一致的。另外,对三株转基因植株的后代幼苗进行胭脂碱检测,表明α'亚基基因在转基因烟草中稳定遗传,后代分离比例为3:1,符合孟德尔遗传规律,故α亚基基因是以单位点的形式整合到受体的染色体上,符合一对因子遗传的规律。

A gene encoding a-subunit of β-eonglyeinin,a seed storage protein of soybean,was transferred into tobacco cells on dis-armed plasmid of Aqrobacterium tumefaciens.Transformed cells were regenerated into whole plants.The presence of a'-subunit gene in regenerated plants was confirmed by southern hybridization ana- lysis.Enzyme-linked immunosorbent assay demostrated that the a'-subunit gene had expressed in tobacco plants and the expressed product accumulated only in tobacco seeds,but not in leaves and stems.Nopaline detection showed that the progenies derived from 3 transgenic tobacco plants inherited the a'-subunit gene in a Mendelian manner.