Characterization of Autographa californica Multiple Nucleopolyhedrovirus (AcMNPV) of Different Isolation

• Main Authors: Wei Kun-Pao, 韋焜寳
• Other Authors: Lee Song-Tay
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/79420215397744676017

碩士 === 南台科技大學 === 生物科技系 === 97 === Using cell biology and molecular biology and proteomic technique, we characterized and compared three Autographa californica multiple nucleopolyhedrovirus (AcMNPV) isolates AcMNPV-TW4 from Taiwan and two other strains AcMNPV-E2 and AcMNPV-C6 from other countries. In this study, the infected Sf-9 cells with these three isolates were examined and compared 3 parameters, CPEs (cytopathic effect); titers of budded virus; and yields of OBs (occussion bodies). In bioassay, the larvae of diamond back moth (plutella xylostella) were infected with three viruses, the larvae mortalifies were counted and compared. In molecular analysis, the identifies of polyhedrin genes and RFLP of the viral genomes were studies. Finally, we explored the proteomic profiles of occlusion body and budding virus of AcMNPV isolates by using 1-D or 2-D electrophoresis. The isolates AcMNPV-TW4 infected cells showed CPEs earlier than the other two isolates. The cell lysis and occlusion bodies releasing of both AcMNPV-TW4 and AcMNPV-C6 infected Sf-9 cells were faster than those of AcMNPV-E2 infected cells. The survival rate of infected cells and the titers of budding viral progeny did not show significant difference among these isolates. However, the occlusion bodies produced by the AcMNPV-TW4 infected cells were more than those by the other two AcMNPVs infected cells. The highest mortality of the infected AcMNPV-TW4 diamond back moth larave were found. The RFLP profifed of HindIII、EcoRI、BamHI and XhoI showed significant difference between AcMNPV-C6 and the other two isolates. The differences also could be detected between AcMNPV-E2 and AcMNPV-TW4. The profiles of polyhedral proteins of AcMNPV-C6 and AcMNPV-TW4 by 1-D electrophoresis were found a significant different in polyhedral protein contents. The amounts of four proteins of budding viruses exhibited a significant difference between AcMNPV-C6 and AcMNPV-TW4 by 2-D electrophoresis, three proteins from AcMNPV-TW4 budding virus were more amount than those from AcMNPV-C6 budding virus, while the last one of AcMNPV-C6 budding virus was more amount. These results showed the discrepancy exists on the different isolated of a certain virus. However, which gene or genes actually contributed to the discrepancy, it is a worth issue for further studies.