全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

PRRS病毒E基因的克隆及表达载体的构建

DOI: 10.3321/j.issn:1000-7091.2006.01.007, PP. 27-30

Keywords: PRRSV,基因克隆,原核表达,真核表达

Full-Text   Cite this paper   Add to My Lib

Abstract:

分别以质粒pBV221和pPIC9K为表达载体,成功构建了PRRS病毒E基因克隆载体,转化后经抗性筛选、PCR扩增、双酶切鉴定,确认得到了含有目的基因的阳性克隆。对含有E蛋白原核表达载体的阳性克隆进行诱导表达,经过酶联免疫鉴定,结果显示外源蛋白在宿主菌中有表达,成功实现了在大肠杆菌细胞中的克隆。本研究成功构建了PRRSVBJ-4毒株E基因的原核表达载体和真核表达载体,为基因工程疫苗的研制奠定了基础。

References

[1]  Mengeling W L,Lager K M,Vorwald A C.Clinical effects of porcine reproductive and respiratory syndrome virus on pigs during the early postnatal interval[J].Am J Vet Res,1998,59(1):52-55.
[2]  乔传玲,郑厚旌,甘孟侯,等.反转录聚合酶反应检测猪繁殖与呼吸综合征病毒的研究[J].畜牧兽医学报,2000,31(3):262-266.
[3]  Pringle C R.Virus Taxonomy 1997-Proceedings of the Excutive Committee of the ICTV [J].ArchVirol,1997,142 (8):1727-1733.
[4]  Sur J H,Doster A R,Osorio F A.Apoptosis induced in vivo during acute infection by porcine reproductive and respiratory syndrome virus[J].Vet Pathol,1998,35(6):506-514.
[5]  奥斯伯F,布伦特R,金斯敦R E.精编分子生物学实验指南[M].颜子颖等,译(第3版).北京:科学出版社,1999.
[6]  Sambrook J,Russell D W.Moecular cloning:A Laboratory Manual 3 rd ed[M].New York:Cold Spring Harbor Laboratory,2001.8:40-55.
[7]  Wensvoort G,Tepstra C,Pol J M A,et al.Mystery swine disease in the Netherlands:the isolation of Lelystad virus [J].Veterinary Quarterly,1991,13:121-130.
[8]  Terpstra C,Wensvoort G,Pol J M A.Experimental reproduction of porcine epidemic abortion and respiratory syndrome(mystery swine disease)by infection with Lelystad virus [J].Vet Q,1991,13:131-136.
[9]  郭宝清,陈章水,刘文兴,等.从疑似PRRS流产胎儿分离PRRSV的研究[J].中国畜禽传染病,1996,18(2):1-4.
[10]  杨汉春,管山红,尹晓敏,等.猪繁殖与呼吸综合征病毒的分离与鉴定[J].中国兽医杂志,1997,23(11):3-5.
[11]  Meulenberg J J M,Besten A P,de Kluyver E P,et al.Characterization of proteins encoded by ORFs 2 to 7 of Lelystad virus [J].Virology,1995,206:155-163
[12]  Delputte P L,Vanderheijden N,Nauwynck H J,et al.Involvement of the matrix protein in attachment of porcine reproductive and respiratory syndrome virus to a heparinlike receptor on porcine alveolar macrophaGes [J].J Virol,2002,76(9):4312-4320.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133