一种检测转1Dx5基因小麦植株的新方法

目的:建立一种有效区分Glu-1D等位基因的Multiplex-PCR体系,快速检测转1Dx5小麦植株。方法:根据1Dx5和1Dx2亚基基因的区别设计特异的PCR引物,通过多重PCR技术检测花粉管通道法转化1Dx5核心片段的转基因T1代植株。结果:Multiplex-PCR能够在转基因T1代材料中扩增Glu-1D等位基因的特征条带,分别为343bp、320bp的1Dx5基因特异片段以及361bp的1Dx2基因特异片段,与预期结果一致。结论:该技术能够检测多个靶基因,有效地区分转基因Glu-1D上的等位基因,证实外源1Dx5基因已整合到受体基因组中,对检测基因组庞大、外源基因序列GC含量高且与内源基因同源性高的转基因小麦十分有效。

An New Approach of Detection of 1Dx5 Transgenic Wheat

Objective:To establish an effective system distinction between Glu-1D allele Multiplex-PCR system.Method:From the differences between 1Dx5 and 1Dx2 gene,the PCR specific primers were designed.The transgenic 1Dx5 T1 plants were detected by multiplex PCR,were obtained from ORF of 1Dx5 gene was transformed into Xindong-26 via pollen tube pathway.Result:the target band of exogenous gene was amplified in T1 generation of transgenic material,the size of amplified products were 343bp,320bp of 1Dx5,and 361bp of 1Dx2.Conclusion:This technique minght effectively distinguish between the alleles Glu-1D by detected multiple target genes,and confirmed the foreign 1Dx5 gene was integrated into the recipient genome.as well as,the technique is very effective for the detection of large foreign gene sequences of high GC content and with high homology to endogenous genes in transgenic wheat.