猪O型口蹄疫病毒重组结构蛋白的纯化及间接ELISA方法的建立

Abstract

Abstract: The complete genes encoding the structural protein of foot and mouth disease virus (FMDV)(VP0,VP1 and VP3) were subcloned into expression vector SUMO,resulting in the fusion expression plasmid SUMO-VP0,SUMO-VP1 and SUMO-VP3,and then transformed into E.coli BL21(DE3) and induced by IPTG,the fusion proteins were expressed at high level.Western blotting was performed to confirm that the expressed fusion proteins could specifically react with antiserum against FMDV.The expressed fusion proteins were purified by affinity chromatography. Based on the fusion protein further purification,novel indirect ELISA (SUMO-VP0-ELISA,SUMO-VP1-ELISA,SUMO-VP3-ELISA) were developed to detect FMDV antibodies in swine by detecting 200 serum samples,indicating that the indirect SUMO-VP0-ELISA,SUMO-VP1-ELISA,SUMO-VP3-ELISA were specific and sensitive.

Key words: foot and mouth disease virus serotype O in swine; recombinant structural protein; SUMO; purification; indirect ELISA

CLC Number:

S852.4^＋3

SONG Ni, WANG Feng-xue, SUN Na, SHI Xin-chuan, WEN Yong-jun, WU Hua. Purification of the Recombinant Structural Protein of Foot and Mouth Disease Virus Serotype O in Swine and Establishment of the Indirect ELISA[J]. , 2013, 40(12): 20-26.

References

1 韦平,秦爱建.重要动物病毒分子生物学[M].北京:科学出版社,2008.
2 殷震,刘景华.动物病毒学[M].第2版.北京:科学出版社,1997.
3 廖德芳,信爱国,高华峰,等.O型口蹄疫病毒P1-2A基因重组杆状病毒的构建及其表达[J].中国畜牧兽医,2011,38(10):81~85.
4 Baker R T.Protein expression using ubiquitin fusion and cleavage[J].Curr Opin Biotechnol,1996,7:541~546.
5 Baker R T,Catanzariti A M,Karunasekara Y,et al.Using deubiquitylating enzymes as research tools[J].Method Enzymol,2005,398:540~554.
6 Catanzariti A M,Soboleva T A,Jans D A,et al.An efficient system for high-level expression and easy purification of authentic recombinant proteins[J].Protein Sci,2004,13:1331~1339.
7 Chatterjee D K,Esposito D.Enhanced soluble protein expression using two new fusion tags[J].Protein Expr Purif,2006,46:122~129.
8 Curry S,Fry E,Blakemore W,et al.Dissecting the roles of VP0 cleavage and RNA packaging in picornavirus capsid stabilization:The structure of empty capsids of foot-and-mouth disease virus[J].Virol,1997,71(12):9743~9752.
9 Domingo E,Mateu M G,Escarmis C,et al.Molecular evolution of aphthoviruses[J].Virus Genes,1996,11(2~3):197~207.
10 Freiberg B,Hohlich B,Haas B,et al.Type-independent detection of foot-and-mouth disease virus by monoclonal antibodies that bind to amino-terminal residues of capsid protein VP2[J].J Virol Methods,2001,92(2):199~205.
11 Hondred D,Walker J M,Mathews D E,et al.Use of ubiquitin fusions to augment protein expression in transgenic plants[J].Plant Physiol,1999,119:713~724.
12 Lea S,Abu-Ghazaleh R,Blakemore W,et al.Structural comparison of two strains of foot-and-mouth disease virus subtype O1 and a laboratory antigenic variant,G67[J].Structure,1995,3(6):571~580.
13 Nomine Y,Ristriani T,Laurent C,et al.Formation of soluble inclusion bodies by HPV e6 oncoprotein fused to maltose-binding protein[J].Protein Expr Purif,2001,23:22~32.
14 Vasquez D,Denoya C D,Latorre J L,et al.Structure of foot-and-mouth disease virus capsid[J].Virology,1979,97:195~200.
15 Waugh D S.Making the most of affinity tags[J].Trends Biotechnol,2005,23:316~320.

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Purification of the Recombinant Structural Protein of Foot and Mouth Disease Virus Serotype O in Swine and Establishment of the Indirect ELISA

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