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目的 :获得纯化抗原用于制备CYP2B6多克隆抗体。方法 :PCR扩增目的基因片段 ,亚克隆入融合蛋白表达载体pGEX - 3b ,构建了重组质粒pGEX/ 2B6。然后将该重组质粒转化大肠杆菌DH5α ,IPTG诱导表达 ,SDS -PAGE分离 ,获纯化融合蛋白GST - 2B6。用GST - 2B6免疫BALB/C小鼠 ,自腹水中获取CYP2B6多克隆抗体 (2B6pAb)。结果 :融合蛋白GST - 2B6 (CYP2B6 2 0 2～ 35 2aa) ,并获其特异的多克隆抗体。结论 :通过构建融合蛋白重组表达质粒pGEX/ 2B6 ,用获得的初步纯化融合蛋白GST - 2B6制备了特异的2B6pAb
Objective: To obtain purified polyclonal antibodies for CYP2B6. Methods: The target gene was amplified by PCR and subcloned into the fusion protein expression vector pGEX - 3b to construct the recombinant plasmid pGEX / 2B6. Then, the recombinant plasmid was transformed into E. coli DH5α, induced by IPTG, and purified by SDS-PAGE. The fusion protein GST - 2B6 was purified. BALB / C mice were immunized with GST - 2B6 and the CYP2B6 polyclonal antibody (2B6pAb) was obtained from ascites. Results: The fusion protein GST - 2B6 (CYP2B6202 ~ 35 2aa) was obtained and its specific polyclonal antibody was obtained. Conclusion: The constructed recombinant fusion protein pGEX / 2B6 was used to prepare the specific 2B6pAb