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鸡贫血病毒VP1基因在大肠杆菌中的高效表达及其生物学特性的初步分析

High Expression of Chicken Anaemia Virus VP1 Gene in E.coli and Analysis of Its Biological Characteristics

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【作者】 刘岳龙秦爱建金文杰叶建强吉荣崔治中段玉友

【Author】 LIU Yue long,QIN Ai jian,JIN Wen jie,YE Jian qiang, JI Rong,CUI Zhi zhong,DUAN Yu you (The Animal Science and Veterinary College,Yangzhou University,Yangzhou 225009,China)

【机构】 扬州大学畜牧兽医学院扬州大学畜牧兽医学院 江苏扬州225009江苏扬州225009江苏扬

【摘要】 将鸡贫血病毒 (CAV)VP1蛋白 44 9个氨基酸中的 42 4个氨基酸 (占 95 % )编码序列分二段 ,分别克隆入表达性载体 pGEX - 5X - 3,在大肠杆菌以GST融合蛋白的形式获得了高效表达。以表达产物免疫小鼠 4次 ,制备了抗CAV的多克隆抗体。通过对CAV感染的MSB1细胞做间接免疫荧光试验 (IFA) ,结果为阳性。这表明表达物保留了天然VP1相关的抗原性。用此抗CAV抗体和所建立的IFA方法对临床CAV凝似病料成功地进行了实验室诊断。

【Abstract】 The sequence encoding 424 of 449 amino acids(aa)of CAV VP1(accounting for 95 per cent)were divided into two fragments,and were separately inserted into pGEX-5X-3 vector.They expressed in the form of GST fusion protein in E.coli. at high level,respectively.The anti-CAV polyclonal antibody was produced in mice immunized four times with the fusion protein,and its specificity was identified positive on CAV-infected MSB1 cells by indirect immunofluorescent assay(IFA).This result showed that the expressed fusion protein retained some antigenicities of the natural VP1.The anti-CAV polyclonal antibody is successfully used in clinical diagnosis of CAV by IFA.

【基金】 江苏省科委自然科学基金 ;扬州大学科研基金资助
  • 【文献出处】 病毒学报 ,Chinese Journal of Virology , 编辑部邮箱 ,2002年01期
  • 【分类号】S852.65
  • 【被引频次】16
  • 【下载频次】102
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