转基因植物中Bt杀虫蛋白的重组噬菌体辅助检测

Recombinant Phage-aided Detection of Bt Insecticidal Protein Expressed in Transgenic Plants

以LRP和棉花总蛋白为本底蛋白 ,纯化的Bt杀虫蛋白为目标蛋白 ,利用噬菌体展示技术从噬菌体的随机七肽库中筛选与Bt蛋白特异性结合的七肽 .ELISA检测表明经过三轮淘筛过程 ,特异性多肽得到了高度富集 ,其中PH5可与Bt蛋白特异性结合 .将筛选出的PH5作为Bt蛋白的“类抗体”用于抗虫转基因植物的检测 ,由此建立了一种新的检测方法 。

CryIAc protein of Bacillus thuringensis expressed in E. coli transformed with pQEBt was purified by Ni-NTA affinity chromotography. One phage (PH5) that binds specifically to Bt protein was selected from a combinatorial library of random 7-mer peptides fused to minor coat protein (p III) gene of the filamentous coliphage M13 through three cycles of biopanning and ELISA analysis. A new method of recombination phage aided detection of insecticidal Bt protein in transgenic plants was developed using PH5. The phages selected should also be useful in the structure study of functional domains of Bt insecticidal proteins.