ApNPV Molecular Biology And The Characterstic Of Baculoviras Infection

We could get basically pure ApNPV by purifying the ApNPV that had infected tussah pupae with the method we summarized in the process of our experiments. After extracting the genome of ApNPV and constructing the DNA fragments library of HindIII and Sal I , we did partial gene sequencing analysis to establish a foundation for the whole genome sequencing analysis. Through the RAPD analysis of ApNPV, w-BmNPV, and BmNPV, we found that ApNPV had the closest relativeness with CfNPV, ApNPV had farther relativeness with w-BmNPV, and BmNPV had the farthest relativeness with ApNPV.In order to detect the infection conditions of NPV to insect, we transferred GFP to pBacPAK8 and co-infected Sf21 cells with Ac-BacPAK6 to get recombinant Ac-BacPAK6-GFP through plaque purification.We infected BmN and Sf21 cells with single virus as control groups and then mixed different viruses to infect BmN and Sf21 cells respectively, we can get the following conclusions: Through the construction of recombinant viruses with GFP, we can examine the conditions of proliferation of the recombinant viruses in different cells and issues of insects; ApNPV can't be able to infect BmN and Sf21 cells. When its DNA is transferred into the cells solely, the cells will show the symptom of apoptosis, which is the manifestation of unaffinity between virus and cell; When accompanied by affinitive viruses, AcMNPV and BmNPV can replicate in BmN and Sf21 cells respectively. As to the appearance of polyhedro-like virions in a few cells, it remains to be testified by constructing recombinant BmNPV with GFP; The virions of AcMNPV and BmNPV can get into BmN and Sf21 cells and then release DNAs. This has not been noticed in previous experiments; The partial homologous analyses of ApNPV between other viruses show that it has closer relativeness with CfNPV than with BmNPV and AcMNPV. Through the analysis of infection, we may conclude that those two viruses which have closer relativeness with each other can infect special cells cooperatively. This may afford valuable information to the research of the expansion of host range.