Scientia Agricultura Sinica ›› 2008, Vol. 41 ›› Issue (7): 2154-2161 .doi: 10.3864/j.issn.0578-1752.2008.07.038

• VETERINARY SCIENCE • Previous Articles     Next Articles

Clone and expression of broiler HSP70 gene in E. coli and preparation of HSP70 monoclonal antibody

  

  1. 南京农业大学动物医学院
  • Received:2006-06-05 Revised:2007-08-15 Online:2008-07-10 Published:2008-07-10

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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

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