论文部分内容阅读
用苏云金杆菌菌株 HD- 1和 HD- 73分别发酵培养制备了孢晶混合物 ,孢晶混合物经分离、纯化、电泳得到 Cry IA、Cry IA( c)杀虫晶体蛋白。用 Cry IA和 Cry IA( c)分别免疫家兔制备了两种多抗。用 Cry IA免疫小鼠 ,经细胞杂交、融合、克隆后筛选出 4个单克隆株系 A4F5F1 1、B4E6C7、B4E6D8、B4F5G1 1。用 Cry IA( c)多抗包被 ,B4E6D8上清液作为夹心抗体成功地建立了双抗夹心 EL ISA,并检测了中棉所 30、NUCOTN33B蕾期叶片中毒蛋白的含量 ,结果分别为 2 3.4ng·g- 1FW、2 4 .7ng·g- 1FW。
The spore mixture was prepared by fermentation of Bacillus thuringiensis strains HD-1 and HD-73 respectively. The spore-crystal mixture was isolated, purified and electrophoresed to obtain Cry IA and Cry IA (c) insecticidal crystal proteins. Two kinds of polyclonal antibodies were prepared by immunizing rabbits with Cry IA and Cry IA (c) respectively. The mice were immunized with Cry IA, and four monoclonal strains A4F5F1 1, B4E6C7, B4E6D8 and B4F5G1 1 were selected by cell hybridization, fusion and cloning. The double antibody sandwich EL ISA was successfully constructed using Cry IA (c) polyclonal antibody and B4E6D8 supernatant as the sandwich antibody. The content of poisoned protein in leaves of Zhongmian 30 and NUCOTN33B was detected, which were respectively 2 3.4 ng · g-1FW, 24.7ng · g-1FW.