关键词: 热休克蛋白70, 原核表达, 单克隆抗体制备, 肉鸡

Abstract: The mRNA of broiler HSP70 gene was amplified by RT-PCR and the production was cloned into the PGEM-T Easy vector and the expression vector pET-28a（+）. The recombinant expression vector was transformed into E. coli BL-21 and induced by IPTG to express. The recombinant broiler HSP70 was purified to prepare the polyclonal antibody and the immunel activity of recombinant broiler HSP70 was proved by western-blot. The recombinant broiler HSP70 was used to immunize BALB/ c mice and two hyridoma cell lines secreting monoclonal antibody of broiler HSP70 were established by hybridoma technique. One of the high-specific monoclonal antibody for broiler HSP70 was selected by western-blot and named as BH70-1. The immunohistochemistry results showed that the monoclonal antibody BH70-1 has high affinity with broiler tissues. The results suggest that the prepared monoclonal antibody BH70-1 is effective.

Key words: Heat shock protein 70, Expression in E.coli, Monoclonal antibody, Western-blot, Immunohistochemistry, Broiler