摘要
为真核表达梅花鹿干扰素β1(IFN-β1)并检测其抗牛病毒性腹泻病毒(BVDV)活性,本研究以梅花鹿肝脏基因组为模板,通过PCR扩增出IFN-β1的编码区序列,克隆于真核表达载体pVAX1中,构建重组真核表达质粒pVAX1-IFNβ1,并转染MDBK细胞进行表达。结果显示,将pVAX1-IFNβ1转染MDBK细胞后经western blot和间接免疫荧光法检测和鉴定证实,转染的重组质粒能够在MDBK细胞中正确表达目的蛋白,表达的重组蛋白约为25 ku。此外,采用细胞病变抑制法检测表明转染后重组细胞表达的IFN-β1具有抗BVDV活性。本研究为梅花鹿IFN-β1蛋白抗病毒分子机制的研究及抗病毒药物的开发奠定了基础。
In order to express Sika deer interferon betal （IFN-131） and detect the antioxidant activity of bovine viral diarrhea virus （BVDV）, the IFN-β1 gene was amplified by PCR from genome DNA extracted from liver of sika deer liver. Then, the IFN-β1 gene was cloned into pVAX1 vector to construct recombinant plasmid of pVAXI-IFNβ1. The pVAXI-IFNβ1 was transfected to Madin-Darby bovine kidney （MDBK） cells. The results show protein was expressed correctly in MDBK cells transfected with pVAXI-IFNβ1 by western blot and indirect immunofluorescence assay. The recombinant proteins was about 25 ku. MDBK cells transfeeted with pVAX1-IFNβ1 had anti-BVDV activity by cytopathie inhibition method. The study provided a basis for antiviral mechanism and antiviral agent development.
作者
刘千辉
李哲
苏凤艳
曾范利
王全凯
LIU Qian-hui LI Zhe SU Feng-yan ZENG Fan-li WANG Quan-kai(Chinese Medicinal Materials College, Jiliri Agricultural University, Changchun 130118, China Sika Deer production and application laboratory, Jilin Agricultural University, Changchun 130118, China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2016年第11期907-910,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
吉林自然科学基金项目(201115194)
国家国际科技合作专项(2011DFA32900)
关键词
梅花鹿
IFN-β1基因
真核表达
sika deer
interferon betal gene
eukaryotic expression
