样品制备对转基因抗虫棉外源Bt蛋白定量检测的影响

采用ELISA(Enzyme-linked immunosorbent assay)方法,以转Bt基因抗虫棉品种中棉所45(ZGK 9822)和鄂抗虫棉1号(GK 19)为研究材料,对叶片的不同组织、同一样品不同的冻存时间和同一样品不同研磨程度的Bt蛋白进行定量检测。结果表明:叶片不同组织Bt蛋白表达量差异较小,叶脉Bt蛋白表达量略高于叶肉和叶柄的Bt蛋白表达量;转基因抗虫棉单位鲜物质质量Bt蛋白含量和单位可溶性总蛋白Bt蛋白含量随着冻存时间的延长而降低,鄂抗虫棉1号单位鲜物质质量Bt蛋白含量冻存90 d样品与冻存30 d、60 d样品差异显著,单位可溶性总蛋白Bt蛋白含量无显著差异;不同研磨程度对转基因抗虫棉单位鲜物质质量Bt蛋白含量和单位可溶性总蛋白Bt蛋白含量都存在影响,其中差异最大的为鄂抗虫棉1号,200目试样比50目试样单位鲜物质质量Bt蛋白含量上升了56%,单位可溶性总蛋白Bt蛋白含量上升了38%。

Effects of Sample Preparation on Quantitative Detection of Bt Protein in Genetically Modified Cotton

In order to evaluate the effects of sample preparation on quantitative detection of Bt protein in genetically modified cotton, several factors, such as different tissue sampled from leaves, different frozen storage time of the same samples and different particle size after grinding, were studied in this work using the transgenic cotton cultivars ZGK 9822 and GK l9 by ELISA. The results showed the content of Bt protein in vein was slightly higher than that in mesophyll and in petiole; Bt protein content per unit fresh mass weight and per unit total soluble protein decreased with the extension of frozen time. Significant differences were observed in Bt protein content per unit fresh mass weight between samples with frozen storage for 90 days and in samples with frozen storage for 60 days or 30 days, while no difference was found in Bt protein content per unit total soluble protein among samples with frozen storage for 30, 60 and 90 days; There was distinct impact on Bt protein content per unit fresh mass weight and per unit total soluble protein in samples with different particle sizes. Compared with samples with 50 mesh in size, Bt protein content per unit fresh mass weight in samples with 200 mesh in size increased by 56% and Bt protein content in unit total soluble protein increased by 38%.