1Dx5+1Dy10品质基因共转化小麦后代的胚乳特异性表达及检测分析  

Endosperm-specific Expression of Transgenic Wheat Co-transferred with High Molecular Weitht Glutenin Subunit(HMW-GS) Genes 1Dx5+1Dy10

作　　者：施农农[1] 王慧中[1] 何光源[2] 李克秀[2] 

机构地区：[1]杭州师范大学生命科学学院生化与分子生物学杭州市重点实验室,杭州310018 [2]华中科技大学中英基因工程与基因组学联合实验室,武汉430074

出　　处：《科技通报》2007年第2期198-201,206,共5页Bulletin of Science and Technology

基　　金：中国博士后基金(2002);浙江省自然科学基金(M303081)资助

摘　　要：采用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)的方法对可育转基因栽培小麦鄂恩1号(EN1)和鄂麦11号(EM11)植株的T1代种子中的谷蛋白亚基成份进行分析,验证种子中是否含有对增强小麦展弹性有显著作用的转基因胚乳特异性表达产物——优质高分子量谷蛋白亚基(HMW-GS)。转基因株D193种子中连锁表达两种亚基基因1DX5和1Dy10,转基因株D185和D186则均表达其中的1Dx5亚基基因,不同基因型受体可能影响转基因的连锁表达。2χ检验表明鄂麦1号、鄂麦11号二种基因型T1代种子中转基因表达蛋白1Dx5和1Dx5+1Dy10产生的表型比均符合预期分离比31∶(P>0.05),两种外源基因在小麦基因组中可能均为单位点整合。表明遗传转化分子育种可以成为改良我国小麦加工品质的有效途径。同时并讨论了SDS-PAGE有效检测转基因在早期世代表达的关键步骤。To detect transgene endosperm-specific expression, wheat transgenic seeds from Chinese wheat cuhivars ENI and EM11 through co-transformation of High Molecular Weight-Glutenin Subunit （HMW-GS）genes 1Dx5＋1Dy10 which were proved to improve the visco-elastic quality of dough in wheat flour were analysed by whole protein extraction and SDS-PAGE method. Two isolated fertile transgenic lines D185 and D186 from genotype EN 1 got expression band of 1Dx5 subunit transgene, one fertile transgenic line D193 from genotype EMIl got co-expression bands of 1Dx5 and 1Dy10 subunit genes. The co-expression of transgenes 1Dx5＋1Dy10 is probably associated with genotype of donor wheat. Chisquare test presented that the observed ratio of expressed phenotype in all three lines matched the expected segregation value of 3：1 （P〉0.05） which shows the integration of single locus of two transgenes respectively. It is also predicted that molecular breeding is the viable approach to improve the quality of Chinese wheat cultivars. The key points to detect transgene expression efficiently in SDS-PAGE procedure were also disccussed.

关 键 词：转基因小麦 SDS—PAGE 1Dx5 1Dy10 Χ^2检验 

分 类 号：Q344.13[生物学—遗传学]