鸡贫血病毒衣壳蛋白的表达纯化和多克隆抗体制备.pdfVIP

第 21 卷 5 期 中 国 病 毒 学 21 （5 ）：477-480 2006 年 9 月 VIROLOGICA SINICA September 2006 鸡贫血病毒衣壳蛋白的表达纯化及多克隆抗体制备 1 2 1 1 1** 王晓艳 ，文 刚 ，高玉龙 ，高宏雷 ，王笑梅 （1.中国农业科学院哈尔滨兽医研究所，黑龙江哈尔滨 150001 ；2. 哈尔滨工业大学海洋学院， 山东威海 264209 ） Expression and Purification of the Capsid Protein of Chicken Anemia Virus and Preparation of Antibody 1 2 1 1 1** WANG Xiao-yan ，WEN Gang ，GAO Yu-long ，GAO Hong-lei ，WANG Xiao-mei (1.Harbin Veterinary Research Institute, CAAS, Harbin 150001, China；2. Harbin Institute of Technology, Weihai 264209, China ) Abstract ：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 SDSand Western blot showed that the target gene was expressed successfully in the form of inclusion body when induced with IPTG. The protein was 2+ then purified by Ni -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 v