转Bt基因棉Bt毒蛋白的ELISA检测方法的研究(英文)  

On the Enzyme linked Immunosorbent Assay of Bacillus thuringiensis Insecticidal Protein Expressed in Transgenic Cotton

作　　者：陈松[1] 吴敬音[2] 程德荣[1] 张荣铣[3] 黄骏麒[1] 

机构地区：[1]江苏省农业科学院经济作物研究所,南京210014 [2]江苏省农业科学院农业遗传生理研究所,南京210014 [3]南京农业大学农学系,210095

出　　处：《棉花学报》1999年第5期259-267,共9页Cotton Science

基　　金：国家计委"八五"攻关项目

摘　　要：利用纯化的Ｂｔ毒素蛋白（６７ｋＤａ）作为标准蛋白和免疫抗原，建立了检测转基因抗虫棉植株内Ｂｔ毒蛋白含量的抗体夹心ＥＬＩＳＡ方法。检测极限１μｇ·Ｌ－ １，线性范围１－ １５μｇ·Ｌ－ １，板内误差Ｃ．Ｖ．＜ ５％ ，板间误差Ｃ．Ｖ．＜ ５％ ，平均样品回收率９４．９４％ 。经稀释度试验等质量控制试验，表明所建立的ＥＬＩＳＡ方法是可靠的，可用于转基因棉花植株中Ｂｔ毒素蛋白含量的测定。An antibody sandwich enzyme linked immunosorbent assay (ELISA) was developed to detect and quantitate the Bacillus thuringiensis (Bt) insecticidal protein expressed in transgenic insect resistant cotton using a purified Bt toxin (67kDa) as standard protein and antigen. The minimal detectable concentration of purified Bt toxin was 1μg·L -1 and the linear assay range of the method was 1μg·L -1 to 15μg·L -1 .Both the errors of intra and interplate(s) were <5%,and the mean recovery rate was 94.94%.It was indicated by quality controal experiments that this ELISA method was stable and reliable ,and it can be used in the detectiomn of Bt toxin content expressed in transgenic cotto.

关 键 词：BT毒蛋白 ELISA 检测 转基因棉 

分 类 号：Q785[生物学—分子生物学] S562.01[农业科学—作物学]