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Virologica Sinica, 21 (5) : 477-480, 2006
Research Article
Expression and Purification of the Capsid Protein of Chicken Anemia Virus and Preparation of Antibody
1.Harbin Veterinary Research Institute, CAAS, Harbin 150001, China
2. Harbin Institute of Technology, Weihai 264209, China
The coding region of capsid protein gene from Chicken Anemia virus(CAV) was amplified from pPro-VP1 by PCR and cloned into pET-30a(+). E.coli BL21(DE3)were transformed by the recombinant plasmid pET30-VP1. Analysis by SDS-PAGE and Western blot showed that the target gene was expressed successfully in the form of inclusion body when induced with IPTG. The protein was then purified by Ni2+-affinity chromatography and used to immunize female Balb/c mice. After three rounds of immunizations, antiserum was collected and used to detect the specificity of recombinant protein. ELISA showed that the titer of antiserum was above 1:12800. Moreover, antiserum reacted specifically with purified recombinant protein in Western blot. This study laid a foundation for the development of CAV diagnostic kit and vaccine.
  Published online: 20 Sep 2006
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