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狂犬病病毒aG株糖蛋白膜外区的原核表达及纯化

Prokaryotic Expression and Purification of Glycoprotein Extracellular Domain of Rabies Virus Strain aG
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摘要 旨在有效检测狂犬病毒抗原及抗体。以狂犬病病毒aG株为模板,反转录扩增糖蛋白膜外区基因,并进行序列测定。将目的片段与表达载体pET-32a(+)分别双酶切、胶回收后相连并转化至Rosseta(DE3)菌株。选取阳性克隆经IPTG诱导表达后,进行SDS-PAGE和Western blotting及质谱鉴定。结果显示,在大肠杆菌中成功表达了糖蛋白膜外区。本研究为建立检测狂犬病毒抗原及抗体的ELISA诊断方法奠定了基础。 The gene of glycoprotein extracellular domain from rabies virus aG strain was amplified by reverse transcriptase-polymerase chain reaction ( RT-PCR ). The product was purified and subcloned into the expression vector pET-32a ( + ). The recombinant plasmid pET-G was transformed into expression strain E. coli Rosetta ( DE3 ) cells and then Isopropyl-J3-D-Thiogalactopyranoside ( IPTG ) was added to induce protein expression. The recombinant protein of glyeoprotein extracellular domain was analyzed by SDS-PAGE, Western blotting and mass spectrometry. Results showed that the recombinant protein had been successfully expressed in E. coll. The study establishes the foundation of ELISA kits to detect the antibodies against rabies virus or antigens of rabies virus.
出处 《生物技术通报》 CAS CSCD 北大核心 2012年第9期114-119,共6页 Biotechnology Bulletin
基金 兰州市生物医药专项(2011-1-61)
关键词 狂犬病病毒 糖蛋白膜外区 原核表达 Rabies virus Glycoprotein extracellular domain Prokaryotic expression
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  • 1Briickner G. Rabies-the role of the World Organisation for animal health in mobilising global control. Vaccine, 2009, 27 ( 51 ) : 7139-7140. 被引量:1
  • 2俞永新主编..狂犬病和狂犬病疫苗[M].北京:中国医药科技出版社,2001:297.
  • 3张伟,吴时友,尹燕博,牛钟相,张秀美.狂犬病诊断基因芯片的建立和检测应用的初步研究[J].中国预防兽医学报,2008,30(2):132-135. 被引量:10
  • 4Zhang KS, Guo JH, Chen HC, et al. Diagnosis and molecular characterization of rabies virus from a buffalo in China : a case report. Virology Journal, 2011, 8 ( 1 ) : 101. 被引量:1
  • 5Benmansour A, Leblois H, Coulon P, et al. Antigenicity of rabies virus glycoprotein. Journal of Virology, 1991, 65 ( 8 ) : 4198-4203. 被引量:1
  • 6Dietzschold B, Wang HH, Rupprecht CE, et al. Induction of protective immunity against rabies by immunization with rabiesvirus ribonucleoprotein. Proc Natl Acad Sci USA, 1987, 84 ( 24 ) : 9165-9169. 被引量:1
  • 7Hu R, Zhang S, Fooks AR, et al. Prevention of rabies virus infection in dogs by a recombinant canine adenovirus type-2 encoding the rabies virus glycoprotein. Microbes Infect, 2006, 8 (4) : 1090-1097. 被引量:1
  • 8Irie T, Kawai A. Further studies on the mechanism of rabies virus neutralization by a viral glycoprotein- specific monoclonal antibody, #1-46-12. Microbiol Immunol, 2005, 49 ( 8 ) : 721-731. 被引量:1
  • 9李伟,张曼夫,张新梅,王树蕙.MBP-狂犬病毒NP融合蛋白在大肠杆菌中的表达及NP的纯化[J].农业生物技术学报,2000,8(1):29-32. 被引量:2
  • 10魏玉荣,易忠,符子华,马素贞,王晓萍,简子健,魏婕,王海烽,朱晶晶.狂犬病病毒SRV_9株N、P、G和L蛋白基因的克隆及表达载体构建[J].新疆农业科学,2009,46(2):354-358. 被引量:8

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