转基因小麦插入突变蛋白的鉴定

Identification of Insertion Mutation Protein of Transgenic Wheat

为研究转基因小麦中插入突变蛋白的编码基因,以转基因小麦B73-6-1和受体小麦L88-6为材料,利用改良方法分离、纯化、回收目的蛋白后进行鉴定。N端测序结果显示,该蛋白N端前五个氨基酸序列(EGEAS)以及部分氨基酸序列与几乎所有高分子量麦谷蛋白亚基(HMW-GS)具有100%的同源性;同时,质谱鉴定结果显示,该蛋白的两个得分最高(96和58)的肽段(GGSFYPGETTPPQQLQQR和IFWGI-PALLKR)与1Dx5亚基的同源性达到100%,初步证明该突变蛋白为新的HMW-GS。这对外源1Dx5基因整合位点的确定和外源基因整合规律的研究提供了更多依据。

To obtain the coding gene of insertion mutation in glutenin of transgenic wheat,the target protein was isolated,purified,recovered and identified by modified methods with material B73-6-1 and L88-6,transgenic wheat and receptor wheat,respectively.It was showed that the first five amino acid sequence（EGEAS） and partial amino acid sequence in the other parts of the target protein had 100% homology with nearly all HMW-GS.Meanwhile,mass spectrometry identification showed that two peptide fragments（GGSFYPGETTPPQQLQQR and IFWGIPALLKR）of the target protein had high score（96 and 58） and 100% homology with 1Dx5.It was preliminarily proved that the mutation in the target protein was a new HMW-GS,which provided basis for the confirmation of integration site of exogenous gene 1Dx5 and the study of integration rule of exogenous gene.