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马立克氏病病毒囊膜糖蛋白B基因原核表达产物单抗的制备
引用本文:韩凌霞,丁家波,陈雷,崔治中. 马立克氏病病毒囊膜糖蛋白B基因原核表达产物单抗的制备[J]. 动物医学进展, 2001, 22(4): 51-53,58
作者姓名:韩凌霞  丁家波  陈雷  崔治中
作者单位:1. 扬州大学畜牧兽医学院,
2. 山东农业大学动物科技学院,
基金项目:国家自然科学基金资助项目 (39870 0 0 8)
摘    要:用大肠杆菌BL21表达了马立克氏病病毒(MDV)强毒GA株的囊膜糖蛋白B(gB)基因,通过SDS-聚丙烯酰胺凝胶龟泳(SDS-PAGE)分离表达蛋白条带,切下并碾碎后作为免疫原制备单克隆抗体。通过酶联免疫吸附试验(ELISA)和间接免疫荧光试验(IFA),得到1株GA株阳性的单克隆抗体杂交瘤细胞株7C8。该单抗腹水的ELISA效价为1:2^12,IFA效价为1:800,与MDV不同致病型毒株CVI988、GA、RB1B、MD11、648A株感染的鸡胚成纤维细胞(CEF)均呈IFA阳性。1:100稀释时与GA感染的CEF在斑点酶联免疫吸附试验(dot-ELISA)中呈阳性。

关 键 词:马立克氏病病毒 gB基因 间接免疫荧光试验 斑点酶联免疫吸附试验 单抗
文章编号:1007-5038(2001)04-0051-03

Preparation of Monoclonal Antibody of Marek''''s Disease Virus (MDV) Glycoprotein B Expressed in E. Coli
HAN Ling xia ,DING Jia bo ,CHEN Lei ,CUI Zhi zhong. Preparation of Monoclonal Antibody of Marek''''s Disease Virus (MDV) Glycoprotein B Expressed in E. Coli[J]. Progress In Veterinary Medicine, 2001, 22(4): 51-53,58
Authors:HAN Ling xia   DING Jia bo   CHEN Lei   CUI Zhi zhong
Affiliation:HAN Ling xia 1,DING Jia bo 1,CHEN Lei 1,CUI Zhi zhong 2
Abstract:Glycoprotein B gene (gB) of virulent MDV GA strain expressed in E.coli was cut off from SDS PAGE ,then inoculated it to BALB/C mice directly to prepare monoclonal antibody. 1 strain of monoclonal hybridoma cells 7C8 was obtained by ELISA and Indirect Fluorescence Assay(IFA). Its ascites fluids titre of ELISA and IFA is 1: 212 and 1:800 respectively. The Mab can also identify CEF infected by MDV through dot ELISA.
Keywords:marek's disease Virus (MDV)  glycoprotein B gene (gB)  indirect fluorescence assay(IFA)  dot ELISA
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