中国动物传染病学报2018，Vol.26Issue(1)：32-36,5.
猪繁殖与呼吸综合征病毒NSP12蛋白的表达与鉴定
PROKARYOTIC EXPRESSION AND IDENTIFICATION OF NON-STRUCTURAL PROTEIN 12 OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS
摘要
Abstract
The non-structural protein12 (NSP12) of Porcine reproductive and respiratory syndrome virus (PRRSV) was amplified and cloned into the pCold-I vector.The recombinant plasmid pCold-Nsp12 was then transformed into E.coli/BL21 cells and induced with 1 mmol/L isopropyl β -D-thiogalactoside (IPTG) at 16℃ for 20 h. The recombinant protein was purified and identified with SDS-PAGE as about 18kDa band and also recognized in Western blotting using PRRSV antisera. Subsequently, the purified recombinant Nsp12 protein was administered into 6-week-old BALB/c mice for production of polyclonal antibodies. The antisera collected from these immunized mice specifically reacted with PRRSV HuN4 strain and NSP12 protein expressed in MARC-145 in IFA and Western blotting. The present study laid a basis for further study on the role of NSP12 in PRRSV infection.关键词
猪繁殖与呼吸综合征病毒/NSP12/原核表达/多抗Key words
Porcine reproductive and respiratory syndrome virus/NSP12/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
张文超,刘欢,童光志,杜雅楠,周艳君,虞凌雪,姜一峰,高飞,黄勤峰,李丽薇,李慧春,陈鹏飞,杨德强..猪繁殖与呼吸综合征病毒NSP12蛋白的表达与鉴定[J].中国动物传染病学报,2018,26(1):32-36,5.基金项目
973计划项目(2014CB542702) （2014CB542702）
国家科技支撑计划课题(2015BAD12B01-1) （2015BAD12B01-1）
国家生猪现代产业技术体系项目(CARS-36) （CARS-36）
