recombinant vp1 protein of fmd virus type o/irn/2010 as an immunogenic peptide expression system

Authors

farahnaz motamedi-sedeh karaj,iran

homayoon mahravani karaj, iran

hoorieh soleimanjahi tehran, iran

lila asadpour gylan, iran

abstract

foot and mouth disease (fmd) is 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. objective: 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. methods: the 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 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 is a good candidate for preparation of peptide vaccine against fmd virus type o/irn/2010 in the next future.

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Journal title:
iranian journal of virology

جلد ۸، شماره ۲، صفحات ۱۲-۱۷

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