转基因小麦T1代中外源基因的检测和分析

该研究采用Multiplex PCR和SDS-PAGE技术对转1 Dx5基因(ORF)T1代小麦进行了检测和分析.结果显示,Multiplex PCR能够扩增出转基因T1代材料中1 Dx2基因和1 Dx5基因的特征片段,表明外源基因已整合到受体基因组中;SDS-PAGE检测到一新的蛋白质亚基X,表明外源基因的插入引起了转基因T1代籽粒中HMW-GS组成的变化.该研究不仅验证了线性基因片段遗传转化策略的可行性,而且印证了普通小麦Glu-1D等位基因的多重PCR分子标记体系的有效性,为培育安全型的转基因作物新种质打下坚实的基础,加快小麦分子育种进程.

Detceting and analysis of foreign gene in T1 generation of transgenic wheat

In order to detect and analyze heritance of Foreign 1Dx5(ORF) in T1 Generation of Transgenic Wheat(The ORF of 1Dx5 was used for transformation of wheat(Xindong-26) via pollen tube pathway).First of all,Characteristic fragment of 1Dx2 and 1Dx5 in transgenic T1 lines by Multiplex-PCR show that foreign gene had been inserted into the genome of wheat cultivar;secondly,a novel protein X was tested in HMW-GS compositions by SDS-PAGE,that may be ascribed to insertion of foreign gene.To sum up,this GM strategy of linear gene is effective and feasible and the PCR-base marker system is available to distinguish between ordinary wheat Glu-1D alleles,which provides a solid foundation to speed up the process of molecular breeding of wheat.