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。
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.
出处
《病毒学报》
CAS
CSCD
北大核心
2002年第1期61-65,共5页
Chinese Journal of Virology
基金
江苏省科委自然科学基金
扬州大学科研基金资助
