摘要
将ＰＲＲＳＶＣＨ１ａ株Ｎ基因用ＥｃｏＲＩ和ＰｓｔＩ双酶切从重组质粒ｐＵＣ１８ＯＲＦ７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ｐＢＶ２２０ＯＲＦ７。转化了ｐＢＶ２２０ＯＲＦ７的大肠杆菌ＪＭ８３经诱导培养后，用ＳＤＳＰＡＧＥ和Ｗｅｓｔｅｒｎｂｌｏｔ检测表达产物。结果表明ＰＲＲＳＶＣＨ１ａ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１５３％。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.Its 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.
出处
《中国预防兽医学报》
CAS
CSCD
1999年第1期35-37,共3页
Chinese Journal of Preventive Veterinary Medicine
关键词
猪
PRRS病毒
N蛋白
原核表达
Porcine reproductive and respiratory syndrome(PRRS)virus Nucleocapsid(N)protein prokaryotic expression
