The Study Of Autographa Californica Multiple Nucleopolyhedrovirus Pk-1Gene Function And Location

Protein kinase is an essential kinase for virus particles to maintain their activity. Baculovirus AcMNPV in their own reproductive processes in addition to the use of host cell protein kinase, itself also an additional encoding a protein kinase---k-1. pk-1is a conserved gene and it is the least Serine/threonine kinase.it exists in almost baculoviridae(except for CuniNPV). It expresses denpend on different baculoviridae. It expresses from12h or48h in AcMNPV and BmNPV, accumulated until which is a very late gene. Instead in ChfuGv and LdMNPV, it expresses from2h or4h arriving at the most high level in the very late infect, so it is a early gene. Linda M. Reily discovered that pk-1have phosporylation in1994. Fan indicated that pk-1is a factor for transcription from very late promoters of poly by temperature sensitive pk-1mutant viral production. This thesis focused on pk-1how to affect viral replication.The first chapter described the baculovirus overview. That include classify and function, focusing on temporal expression patterns of baculovirus genes, the factor involved in the formation of polyhedra. We described some VAPK. At the end, the aims and significance of this thesis was presented.The second chapter represented the materials and methods.The thid chapter described the major results. Analysed pk-1via transfection-infection test. The results show that the absence of pk-1gene can not complete the infection cycle. pk-1reverse mutation can completely rescue the impact due to the lack of pk-1. This indicates that the pk-1gene is a must gene of AcMNPV infection. Real-Time PCR results show that pk-1does not affect DNA replication in the AcMNPV infection. Pk-1deletion affects the normal assembly of the nucleocapsid by electron microscopy sections. Deletion of the AcMNPV pk-1cells can only form a hollow long tubular structures. This implies that pk-1deletion may undermine the process of viral DNA and capsid protein assembly. In addition, PK-1in the viral replication process have a different orientation, in the pre-positioning of infection in the whole cell, in the late mainly localized in the cytoplasm by immunofluorescence This implies that PK-1may be a multi-functional genes play different roles in different infection periods.The fourth chapter summarized and discussed the result of this experiment.In summary, this thesis mainly focused on the fuctional analysis of AcMNPV pk-1which gived a basis on researched pk-1-mediated phosphorylation.