Cloning And Expressing FMDV Non-Structure Protein 3ABC, 3AB, 3A And Establishment Of An Indirect ELISA For Detection Of Antibodies Against 3A Proteins

Foot-and-mouth disease(FMD) is a highly contagious disease of animals which caused by foot-and-mouth disease virus(FMDV). In developing countries,systematic control measure based on vaccination is usually used. Establish an accurate and fast distinction between infected live virus vaccine and injected animals in the detection and diagnosis of FMD control the incidence of livestock quarantine areas are very urgently needed.Infect animals produce antibodies to both the structural and non-structural proteins.Acorroding to the tecnology of the vaccines produce,the structural proteins antibodies are most predominant among the vaccinated animals , the non-structural proteins antibodies little.So using non-structural protein as the antigen to differentiate infectd from vaccined animals is a good method .3ABC gene of foot-and-mouth disease virus(FMDV) was amplified using PCR from FMDV and then cloned into PET-32a expression vector . The recombinant plasmid pET - 32a -3abc was transformed into BL21 , and the purpose of protein was induced by IPTG in BL 21, the PET -32a -3ABC protein was examined and by SDS-PAGE .The result showed that Recombinant vector was expressed in Ecoli successfully . the 3AB,3A gene fragment amplified by RT-PCR from the template of PGEM-T -3ABC ,and then cloned into PET-28a and PET-41a expression vector .The recombinant plasmids were transformed into BL21 , and the purpose of proteins were induced by IPTG in BL21, the proteins were examined by SDS-PAGE and Western blot .The result showed that recombinant proteins were expressed in Ecoli successfully .Using purified production of 3A,an indirect ELISA for detection of FMDV antibodies was developed and its optimal recation conditions wer determined. This research provided technical supportfor ELISA methord to distinguish the infection from vaccination.