Detection of foot-and-mouth virus antibodies using a purified protein from the high-level expression of codon-optimized, foot-and-mouth disease virus complex epitopes in Escherichia coli.
A codon optimized DNA sequence coding for foot-and-mouth disease virus (FMDV) capsid protein complex epitopes of VP1 amino acid residues 21-40, 135-160, and 200-213 was genetically fused to the C-terminal end of a glutathione-S-transferase (GST) gene in pGEX-6P-1 vector with the synonymous codons preferred by Escherichia coli . The gene was synthesized using PCR and subsequently expressed in E. coli producing an intracellular, soluble fusion protein that retained antigenicity associated with FMDV antibodies by western blot analysis. The chimera was purified from bacterial lysates by affinity chromatography and could be used in ELISA tests for antibodies against FMDV.
Centre for Molecular Microbiology, Institute of Microbiology, Chinese Academy of Sciences, 13 Zhongguancun Beiyitiao, Beijing 100080, PR China. songhouhui@yahoo.com.cn
Houhui
H
Song
Weihuan
W
Fang
Zhiliang
Z
Wang
Dongxia
D
Zheng
Jian
J
Du
Hong
H
Li
Yong
Y
Li
Bingsheng
B
Qiu
D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D002417:Cattle; D003062:Codon / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005536:Foot-and-Mouth Disease / Q000097:blood / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D012756:Sheep; D013552:Swine
D000906:Antibodies; D036022:Capsid Proteins; D003062:Codon; D000939:Epitopes; D011993:Recombinant Fusion Proteins