title: [Expression and utilization of 3AB nonstructural protein of foot-and-mouth disease virus in Escherichia coli].
abstract: To develop a sensitive and specific ELISA for detection of antibodies to the nonstructural protein of FMDV. We cloned and expressed FMDV nonstructural protein 3AB in Escherichia coli expression system. The recombinant protein 3AB was purified with Ni-NTA HisBind Resins and characterized by Western blotting. An indirect ELISA based on purified protein 3AB as a coating antigen was established. The specificity and sensitivity of this assay were evaluated by comparison with a commercial 3ABC-ELISA kit in detecion of serum samples. The results showed that the recombinant protein 3AB was expressed as a formation of inclusion bodies in Escherichia coli. The purified protein could specificially react with FMDV infection antibodies in Western blotting assay, but no reaction with the immune antibodies induced with vaccine. Two assays were no significant differences in specificity and sensitivity for detection of field samples (P>0.05). Therefore, we speculated that the recombinant protein 3AB is a promising molecular marker, which may effectively differentiate FMD-infected from vaccinated animals in a herd.
authors.affiliation: Key Laboratory ofAnimal Virology of Minister of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
authors.forename: Junjun
authors.initials: J
authors.lastname: Shao
authors.forename: Huiyun
authors.initials: H
authors.lastname: Chang
authors.forename: Tong
authors.initials: T
authors.lastname: Lin
authors.forename: Guozheng
authors.initials: G
authors.lastname: Cong
authors.forename: Junzheng
authors.initials: J
authors.lastname: Du
authors.forename: Shandian
authors.initials: S
authors.lastname: Gao
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000276:immunology; D029544:Foot-and-Mouth Disease Virus / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D005822:Genetic Vectors / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology
chemical_list: C109754:3ABC protein, virus; D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins