样品制备对转基因抗虫棉外源Bt蛋白定量检测的影响.pdfVIP

棉 花 学报 Cotton Science 20 15袁27 2 冤院184―188 品制备对转基因抗虫棉外源Bt 蛋白 定量检测的影响 丽娟 刘文娟 张富丽 王东 尹全 雷绍荣 郭灵安 宋君* 四川省农业科学院分析测试中心袁成都6 10066 冤 摘要院采用ELISA Enzyme-linked immunosorbent assay冤方法袁以转Bt 基因抗虫棉品种中棉所45 ZGK 9822 冤 和鄂抗虫棉1 号 GK 19冤为研究材料袁对叶片的不同组织尧同一样品不同的冻存时间和同一样品不同研磨程度 的Bt 蛋白进行定量检测遥 结果表明院叶片不同组织Bt 蛋白表达量差异较小袁叶脉Bt 蛋白表达量略高于叶肉 和叶柄的Bt 蛋白表达量曰 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袁鄂抗虫棉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 尧60 d 样品差异 显 袁单位可溶性总蛋白Bt 蛋白含量无显 差异曰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袁其中差异最大的为鄂抗虫棉1 号袁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% 袁单位可溶性总蛋白Bt 蛋白含量上升了38%遥 关键词院转基因抗虫棉曰样品制备曰Bt 蛋白含量曰定量检测曰ELISA 中图分类号院S562.035.3 文献标志码院A 文章编号院1002-7807 20 15冤02-0 184-05 院10.11963/issn.1002-7807.201502012 Chang Lij uan, Liu Wenjuan, Zhang Fuli, Wang Dong, Yin Quan, Lei Shaorong, Guo Lingan, Song Jun* ( 610066, ) 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 differ- ent 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