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Objective: human respiratory syncytial virus (Human respiratory syncytial virus, RSV) is widely distributed around the world, is the most important viral pathogen of infants and young children, the elderly and immunocompromised patients of severe lower respiratory tract infection. RSV mainly in and antigen RSV belongs to the paramyxovirus Division lung virus genus, gene group to 15.2 Kb of single-stranded negative chain RNA encoding 11 kinds of proteins, including adhesion protein (Glycoprotein, G) and fusion protein (Fusion protein, F), matrix proteins (Matrix protein, M) skeleton protein of RSV. From formalin-inactivated vaccine has been developed by various forms of RSV vaccine, but due to issues such as safety and immunogenicity, as yet not been able to for the prevention of RSV infection, the vaccine will be available. Virus-like particle vaccine as a new form of a vaccine is widely used for a variety of viral vaccine development and the ideal immune effects and protective effects. So far, it is for the preparation conditions of the RSV virus-like particles (Virus-like particles, VLPs) is still in the exploratory stage, only about RSV VLPs found in a U.S. patent application in 2008, but there are low yield, preparation difficult and no immune effects and immune protection of research. This study is intended for use in the first generation of non-replicating adenovirus vector (First generation replication deficient recombinant adenoviral vector FGAd) and baculovirus expression vectors were constructed to express RSV F, G, M replication-defective recombinant adenovirus and recombinant rod The virus, in order to further clarify RSV VLPs formation and budding mechanisms as well as RSV VLPs experimental basis. : The reference GenBank accession RSV A subtype Long strain M, G protein gene sequence, the mammalian cell codon bias, codon optimization of the M and G genes were cloned into the shuttle vector pShuttle-CMV, and then by The homologous recombination adenovirus plasmids were transfected into 293 cells, obtained the recombinant adenovirus FGAd-Gsyn, and FGAd-Msyn Western blot identification purposes gene expression. Reference GenBank accession Isoform Long strain of RSV A F, G and M protein gene sequence, in accordance with the the insect cells codon bias, F, G and M gene codon optimized, then were cloned into to shuttle vector pFast-Bac I transfected the seat after recombinant baculovirus plasmid were transfected sf9 cells, obtained recombinant the baculovirus, rBac-Fopt was, rBac-Mopt and rBac-Gopt, Western-blot identification of target gene expression. Results: The recombinant adenovirus DNA molecules transfected 293 cells, the cells in about seven days, swelling round shrinkage typical cytopathic effect (Cytopathic effect, CPE), Western blot detection to the G and M gene expression, access to non-replicating The to recombinant adenovirus FGAd-Gsyn, and FGAd-Msyn. Recombinant baculovirus plasmid after transfection sf9 cell, 3d cell swelling, enlarged nuclei and nuclear vacuoles lesions Western blot detected the F, G and M gene expression, obtained recombinant the baculovirus, rBac-Fopt was the the rBac-Mopt and rBac-Gopt o Conclusion: The success FGAd-Gsyn, FGAd-Msyn, rBac-Fopt, rBac-Mopt and, rBac-Gopt formation and budding mechanism to clarify the RSV VLPs and RSV VLPs experimental basis.
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