恶性疟原虫MSP1蛋白C-末端19kD蛋白在家蚕杆状病毒表面展示系统中的表达研究

Plasmodium falciparum merozoite surface protein1(MSP1) is one of the most important antigens in Malaria vaccine research.Produced by hydrolysis of the c-Terminal 19kD (MSP119c) fragment is still exist in the merozoite surface after the merozoite enters human red blood cells.research suggests that T-cells and B-cells can be induced by the organism immune response,so it becomes the important candidate antigen locus.In this paper we connect the MSP119c genes to a show caeeier of pFastBac I-gp64,which is built in our laboratory,gp64 genes containing signal peptide sequence and gene of transmembrane sequence.We uccessfully built the eukaryotic for physical size pFastBac I-gp64-MSP119c.Using this recombinant plasmid transformation the DH10Bac Escherichia coli competent cells.After that we got the recombinant Bacmid-gp64-MSP119c silkworm baculovirus genomes.Liposome-mediated transfection of silkworm BmN cells,after assembling formation appers recombinant baculovirus BmNPV-gp64-MSP119c and copy the amplification in BmN cells.The BmNPV-gp64-MSP119c recombinant virus infection in the middle of the third generation complex vaccination SilkWorm, collects incidence chrysalis after 7 days.Bi-directional electrophoresis and Western Blotting showed that recombinant baculovirus MSP119c protein expressed in Silkworm pupa, proofed that recombinant baculovirus recombinant BmNPV-gp64-MSP119c has successed built.Purification recombinant viral particles and immuning New Zealand male rabbits serum antibody preparation. Antibodies are purified by ProteinA antibody purification columns,ELISA detection of antibody to discover the potency of 1:8,000 or more.the surface of the recombinant viruses can effectively encourage the body to produce antibodies. Study on MSP119c protein of Plasmodium falciparum was successfully shown in Silkworm baculovirus surface, laid the Foundation for a new malaria vaccine preparation.