猪生殖-呼吸道综合征病毒CH-1_株N基因的原核表达  

Expression of the N Gene of Porcine Reproductive and Respiratory Syndrome Virus Strain CH 1a in E.coli

作　　者：仇华吉[1] 童光志[1] 郭宝清 王柳[1] 张绍杰[1] 王玫[1] 蔡雪晖[1] 于力[1] 刘宝全 

机构地区：[1]中国农业科学院哈尔滨兽医研究所生物技术国家重点实验室

出　　处：《中国预防兽医学报》1999年第1期35-37,共3页Chinese Journal of Preventive Veterinary Medicine

摘　　要：将ＰＲＲＳＶＣＨ１ａ株Ｎ基因用ＥｃｏＲＩ和ＰｓｔＩ双酶切从重组质粒ｐＵＣ１８ＯＲＦ７切下后，插入到原核表达载体ｐＢＶ２２０的ＰＲ、ＰＬ启动子下游，得到重组表达质粒ｐＢＶ２２０ＯＲＦ７。转化了ｐＢＶ２２０ＯＲＦ７的大肠杆菌ＪＭ８３经诱导培养后，用ＳＤＳＰＡＧＥ和Ｗｅｓｔｅｒｎｂｌｏｔ检测表达产物。结果表明ＰＲＲＳＶＣＨ１ａ株的Ｎ基因在原核载体上得到高效表达，表达产物占菌体总蛋白的１５３％。表达蛋白可望成为有价值的ＰＲＲＳ诊断抗原。Porcine reproductive and respiratory syndrome(PRRS) is a newly recognized pig disease of economic importance characterized by reproductive failure in sows and increased pre weaning mortality in piglets.Its caused by porcine reproductive and respiratory syndrome virus(PRRSV),a small spherical enveloped RNA virus.PRRSV strain CH 1a was recently isolated in a pig farm in China during an outbreak of PRRS.It has been shown to be North American genotype,and its N gene has been cloned,sequenced,and analyzed elsewhere.The N gene was inserted into downstream of the P L and P S promoters of pBV220 (an expressing plasmid introduced from a Chinese institute).The recombinant plasmid pBV220 ORF7 was thus constructed.Strain JM83 of E.coli.containing pBV220 ORF7 was induced at 42℃ for 4 7hr after incubation with vigorous shaking at 37℃ for 3hr or so.A unique protein band of approximate 15 kDa was characterized in the lysate of the transformed bacteria by SDS PAGE electrophoresis.The expression level was up to 15 3% of the total bacterial proteins.Western blot analysis showed the expressed protein was specific to antisera against PRRSV strain Ch 1a.The recombinant N protein is potentially valuable antigen for serological tests and immunoprophlaxis of PRRS.

关 键 词：猪 PRRS病毒 N蛋白 原核表达 

分 类 号：S852.651[农业科学—基础兽医学]