重组抗原CP4-EPSPS的克隆、表达和纯化

Cloning,expression and purification of recombinant antigen CP4-EPSPS

构建了一个含有CP4-EPSPS外源基因的细菌表达载体并在E.coli BL21(DE3)中的高效表达,通过SDS-PAGE检测发现,目的蛋白完全以可溶性方式存在于细胞破碎上清液中。收集该上清液并进而通过镍柱亲和层析,CP4-EPSPS重组蛋白纯度可达85%以上,利用Brandford法测定蛋白质浓度为0.9 mg/m L。该蛋白表达和纯化体系可为转基因植物、食品等蛋白检测用抗体提供稳定免疫性抗原,同时也可为蛋白标准物质研制提供稳定物质基础。

A bacterial expression vector containing CP4 -EPSPS exogenous gene was constructed and expressed efficiently in E. coli BL21 （ DE3 ）. SDS - PAGE showed that the target protein was completely soluble in the cell supernatant liquor. The supernatant was collected and further purified by nickel column affinity chromatography. The purity of CP4 -EPSPS protein was above 85% , and the protein concentration was 0.9 mg/mL determined by Brandford method. The protein expression and purification system can provide stable immune antigen for the transgenic plants, food and other antibody for protein detection, and also provide a stable material basis for the preparation of protein standard material.