Preparation Of Monoclonal Antibody Against M Protein Of Spring Viremia Of Carp Virus And Preliminary Study About Common Carp Natural Immunity Protein TRIM32

Spring viraemia of carp(Spring viremia of carp, SVC) is a highly contagious acute viral hemorrhagic disease, which could cause high mortality in juvenlie carp(30%-70%). It mainly infects carps, especially the common carp and it was listed as notifiable in International Aquatic Animal Health Code of the Office International des Epizooties(OIE). The initial outbreak of this disease was in European countries caused huge economic losses. However, recently, it has been reported in the USA, Middle East, China, Canada and Brazil. For a clearer understanding of the pathogenesis of SVCV and do the countribution to the prevention. Our study then followed with two aspects.1.The preparation of SVCV/M protein monoclonal antibodyBy PCR amplification of the sequence of spring viremia of carp virus matrix protein(M), the amplified product was cloned into the expression vector pGEX-KG and then get the recombinant plasmid SVCV-M-KG. We immunited 4-week-old BALB/c female mices with purified fusion protein SVCV/M-KG. After the fourth immunization, mice splenocytes were harvested and fused with SP2/0 using polyethylene glycol, and finally we selected one monoclonal antibody 5A1. Indirect immunofluorescence and Western blot show that the antibody can specifically recognize SVCV/M protein, and further analysis showed that the interaction sites of the antibody to the M protein are located between 337 to 522 bp.2. A preliminary study of innate immunity protein TRIM32 of common carpAccording to zebra fish, tilapia and other known TRIM32 gene sequence, we designed random primers to amplify the core fragment of common carp TRIM32 gene, then obtained the whole sequence of TRIM32 by RACE. Biological analysis showed the gene was highly conservative. It has the highest homology with the zebrafish, and phylogenetic analysis showed the same results. Spring viremia of carp virus M protein monoclonal antibody and preliminary research of TRIM32 protein, laiding the foundation for the pathogenic mechanism and control methods of SVCV.