河南农业科学2016，Vol.45Issue(3)：141-143,160,4.DOI:10.15933/j.cnki.1004-3268.2016.03.028
鸭瘟病毒UL6基因的原核表达及多克隆抗体的制备
Prokaryotic Expression and Polyclonal Antibody Preparation of Duck Plague Virus UL6 Gene
摘要
Abstract
To prepare mouse anti-duck UL6 of duck plague virus polyclonal antibodies,a pair of specific primers were designed,UL6 gene was amplified by PCR,the fragment of the UL6 gene was cloned into the prokaryotic expression vector pET-32a( +),the resultant recombinant plasmid pET-UL6 was constructed and transformed into E. coli BL21(DE3). Optimally expressed under the induction of 1. 0 mmol/L IPTG at 37 ℃ for 4 hours. SDS-PAGE analysis showed that the UL6 protein was successfully expressed, and it could be recognized by DPV positive serum. After Westernblot identification,the antiserum against UL6 protein was produced by immunizing Balb/c mouse with recombinant protein. ELISA results showed that the antibodies titer was 1∶16 000. In this research,UL6 gene was successfully expressd in E. coli,the immunogenicity of the polyclonal antibodies was good,and the antiserum could be used for detection of UL6 gene expression.关键词
鸭瘟病毒/UL6基因/原核表达/多克隆抗体Key words
duck plague virus/UL6 gene/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
吴双,马丽丽,朱善元,陆辉..鸭瘟病毒UL6基因的原核表达及多克隆抗体的制备[J].河南农业科学,2016,45(3):141-143,160,4.基金项目
国家自然科学基金项目(31440083) （31440083）
江苏省科技支撑计划---农业部分项目(BE2012366) （BE2012366）
