Recombinant VP1 Protein of FMD Virus Type O/IRN/2010 as an Immunogenic Peptide Expression System
Author(s) -
Farahnaz Motamedi-Sedeh,
Homayoon Mahravani,
Hoorieh Soleimanjahi,
Lila Asadpour,
Kamalodin Shafaei
Publication year - 2014
Publication title -
iranian journal of virology
Language(s) - English
Resource type - Journals
eISSN - 2588-5030
pISSN - 1735-5680
DOI - 10.21859/isv.8.2.3.12
Subject(s) - virology , recombinant dna , polyclonal antibodies , biology , virus , microbiology and biotechnology , antiserum , expression cassette , antibody , gene , vector (molecular biology) , genetics
Background and Aims: Foot and Mouth Disease (FMD) is a highly contagious disease among cloven-hoofed animals. FMD virus has structural and non-structural proteins. Vp1 is the most immunogenic structural peptides of FMD virus, applied for major vaccine studies. The aim of this study was construction of Pet28-VP1 cassette for FMD virus type O/IRN/2010 and expression VP1 peptide as the most immunogenic antigen was the aim of this study. Materials and Methods: FMD virus type O/IRN/2010 was isolated from cattle in Qom, Iran and propagated on IBRS2 and BHK21 cell lines. The VP1 gene was amplified using the specific primer pair and RT-PCR technique. The purified PCR product was sub-cloned into the unique BamHI and Xho1 cloning sites to construct the PTZ57R/T -VP1 cassette. The DH5α strain was transformed with this cassette. The digested VP1 gene was cloned in the digested Pet28 and confirmed using double digestion. Then the Pet28-VP1 construction was transformed in BL21 strain. Results: Expression of VP1 peptide was evaluated by IPTG induction and SDS-PAGE and confirmed by Guinea pig specific polyclonal antibody against FMD virus type O and conjugated rabbit anti Guinea pig antibody- HRP. Also neutralizing antisera titre was protective for vaccinated animals by recombinant VP1 protein. Conclusion: Since the isolation of new FMD virus strains in different geographical locations and expression of VP1 peptide which can be used in emergency and control settings as a recombinant vaccine in the same area. Therefore the Pet28-VP1 cassette was constructed in this study may be a good candidate for preparation of peptide vaccine against FMD virus type O/IRN/2010 in future.
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