The Interaction Between Host Cellular Proteins And Autographa Californica Multiple NucleopolyhedrovirusP40 And P49

Baculovirus is a kind of insect virus with the double chain closed circular DNA genome specially infected arthropods. It has a typical biphasic life cycle, there are two different forms of virus particles BVand ODV in its life cycle. They have different functions in the virus life cycle. In the process of baculovirus life there are lots of protein and protein interactions, including the interactions between viral proteins as well as host and viral protein. Autographa californica multiple nucleopolyhedrovirus （AcMNPV） isthe most studied and most thorough baculovirus and widely used in the study.p40and p49 are two baculovirus core genes.Their products are both located in the nuclear capsid of BV and ODV. Previous studies have shown that P40 interacted with a variety of viral proteins.p49 participates in the regulation of cell apoptosis. This study taking the AcMNPV P40, p49 asbait generespectively, using yeast two hybrid technologyscreening of protein interaction respectively from Spodoptera frugiperda cell line （Sf9） cDNA library and cDNA library infected by AcMNPV for 18 hours,, analyzed the interaction of P40 and P49 with host insect protein, the results are as follows:1, Screening the proteins interacted with P40 from Spodoptera frugiperda cell line （Sf9） cDNA library by yeast two hybrid 17 positive clones were got. Three kinds of cDNA respectively clones encode ribosomal protein RL41, the Arp2/3 p21compoundsand CG12863 similar protein withunkown function; the other four cDNA clones code contain part of the coding sequence of Cathepsin precursor, glyceraldehyde 3-phosphate dehydrogenase, hypothetical protein TcasGA2_TC001491 and hypothetical protein KGM₀1973 homologous proteins.2, Screening the proteins interacted with AcMNPV P40 from cDNA library infected by AcMNPV for 18 hours, seven positive clones contain dysbindin-like protein homologous protein coding sequenceand the moth part of an unknown protein homologous sequences.3, Prokaryotic expression and purification of P40 and Sf RL41, use them as antigen immune rabbit to prepare antiserum respectively. The time course analysis ofP40,whichconfirmedp40 is a late expressed gene.4, The result of Co-Immunoprecipitation analysis showed that the RL41 and P40 coprecipitation, which confirmed there is the interaction between the two proteins.5, The result of Co-Immunoprecipitation analysis showed that the P21 and AcMNPV P40 coprecipitation, which confirmed there is the interaction between the two proteins.6, Screening the proteins interacted with P49 fromSpodoptera frugiperdacell line （Sf9） cDNA library by yeast two hybrid,15 positive clones were obtained.These cDNA clones include part of the coding sequences of inhibin protein 2, serine protease precursor, KGM11768, glyceraldehyde 3-phosphate dehydrogenase and ornithine dehydrogenase homologous proteins.