C-terminal region of VP1 of selected foot-and-mouth disease virus serotypes: expression in E. coli and affinity purification.
Foot-and-mouth disease (FMD), one of the most contagious and economically important diseases of farm animals, is caused by a FMD virus (FMDV) which belongs to the family of Picornaviridae. The virus occurs as seven serotypes of which four (A, O, C and Asia 1) are prevalent in India. Immunoprophylaxis supported by precise diagnosis is the prime requirement for achieving the success in controlling the disease. Recently, recombinant DNA technology is gaining importance for the production of cost-effective and safer diagnostics and immunogens. Based on this approach, cDNA of a part of gene for major variable immunogenic region, VP1, of FMDV of four serotypes (A22, O, C and Asia 1) was amplified by PCR and cloned into expression vector. The expression of the 16 K protein gene from the clones was induced with IPTG and analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) and [35S]-methionine labeling. The immunoreactivity of the labeled proteins was assayed by immunoprecipitation with anti-FMDV type-specific sera. Since the proteins contain 6 His residues at the N-terminal end, their affinity purification was carried out using nickel nitrilo-tri-acetic acid (Ni-NTA) agarose matrix. The proteins were found to be immunoreactive and the useful in the FMD diagnosis.
Ratish
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Indian Veterinary Research Institute, Hebbal, Bangalore, India.
Viswanathan
S
S
Suryanarayana
V V
VV
D005537:Aphthovirus / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D002213:Capsid / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D036022:Capsid Proteins; D002846:Chromatography, Affinity / Q000379:methods; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011233:Precipitin Tests; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification; D012703:Serotyping
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C442501:VP1 protein, Foot-and-mouth disease virus