摘要
利用RT-PCR技术扩增了牛病毒性腹泻病毒(BVDV)梅花鹿分离株E0基因,并将其克隆到pMD18-T克隆载体和pET28a原核表达载体中,构建了pMD18-T/E0和pET28a/E0重组子,并进行了测序和表达。表达产物分别用12%SDS-PAGE检测和牛抗BVDV阳性血清进行Western-blotting检测,结果表明,BVDV梅花鹿分离株E0基因的核苷酸序列与匈牙利VEDEVAC株的同源性高达98.6%,与C24V标准株的同源性为84.9%,证明构建的重组子是正确的;BVDV梅花鹿分离株E0基因得到了正确表达。
The E_0 gene of bovine viral diarrhea virus in Sika deer was amplified with RT-PCR,cloned into Cloning vector pMD18-T and prokaryotic expression vector pET28a,sequenced and expressed.Expressed product of the E_0 gene to be tested by SDS-PAGE and Western-blotting.Results showed that the isolated strain,as compared with 2 strains BVDV(VEDEVAC,C_ 24 V),the identities of nucleotide sequences were 98.6% and 84.9%,respectively.The gene E_0 from an isolated strain in Sika deer was expressed correctly.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2005年第4期353-355,共3页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30300256)
吉林省科技发展计划资助项目(20030552-2)
