昆虫细胞表达重组H5亚型禽流感HA蛋白及其免疫原性研究-预防兽医学专业论文.docxVIP

万方数据 万方数据 Abstract H5N1 subtype highly pathogenic avian influenza viruses (AIV) not only cause disaster to poultry industry, but also threaten to human public health security. Immunization of inactive vaccine is one of the effective strategies to prevent and control this disease. The process of screening of vaccine candidate is time-costing and labor-consuming procedures. The production of vaccine is involving live virus and restricted by suppling of chicken eggs. The AIV subunit vaccine based on hemagglutinin (HA) gene can avoid the disadvantages of inactivated vaccine. Hemagglutinin protein which is one of the domainant surface antigens, can elicite antibody in animal and is the primary target protein of development of subunit vaccine. Previously studies showed that the biological activity, antigenicity and immunigenecity of HA protein and Matrix protein were expressed by baculovirus system are similar to natural HA protein in virions. But there are only few reports abou the biological activity, antigenicity and immunigenecity of HA protein were expressed by baculovirus system are similar to natural HA protein in virions The current study adopted baculovirus system to expression of recombinant HA derived from H5 subtype of AIV, and characterized the hemagglutinin activity, probability of assembles of virus-like particles and immunigenecity of the recombinant HA protein. Construction of HA contained clone vector. Two stratigies werte were employed to get HA gene. One of the strategies was cloned HA gene from H5 subtype AIV (Re-6) detection antigen with RT-PCR, and named as L-HA. In another method, the consensus HA gene was selected from the results of aligment of HA gene from GenBank, named as H-HA. The selected HA genes were synthesized after optimization of codon based on codon usage bias of insect cell. The intergrity HA gene (1857 bp) or lack of signal peptide and transmembrane of HA gene (1701 bp) was amplified by PCR with templates of synthesized plasmid(HHA