Expression and purification of recombinant chimeric PvpA-pMGA۱.۲protein from Mycoplasma gallisepticum in Escherichia coli.

BACKGROUND AND ABJECTIVEMycoplasma gallisepticum is the primary agent of chronic respiratory disease in chickens,and infectious sinusitis in turkeys creating important economic losses in poultry industry,which is spread worldwide. pMGA and pvpA are two gene families of Mycoplasmagallisepticum that encode major surface proteins containing pathogenic, antigenic andimmune evasion attributes. The objective of the present study was to express and purify thechimeric PvpA-pMGA۱.۲ recombinant protein from Mycoplasma gallisepticum for designingELISA test.MATERIALS AND METHODSDominant pattern with maximum covering was selected. The codon optimized sequence wascloned into the expression vector pET۳۲a+ and transformed into Escherichia coli strainBL۲۱(DE۳), and the expressed protein was identified by SDS-PAGE.RESULTS AND DISCUSSIONThe results revealed that the recombinant chimeric PvpA-pMGA۱.۲ protein was highlyexpressed post- induction with ۰.۱ mM IPTG after ۱۶ hours incubation at ۳۷˚C and proteinwas purified amount ۱۳۸ mg per liter by affinity Batch formation method. The purifiedrecombinant protein was confirmed by Western blotting.CONCLUSIONThis recombinant chimeric protein could be potentially used for the development ofserodiagnostic test.

کلیدواژه ها:

rPvpA-pMGA۱.۲ ، Mycoplasma gallisepticum ، Fusion protein