Study On ORF82 Function Of Autographa Californica Multiple Nucleopolyhedroviruses

Baculovirus is a kind of double stranded superhelix DNA insect virus.Because baculovirus is harmless to the environment and human beings,it plays an important role in biological control,drug research and development,vaccine production and gene therapy.After the completion of the whole genome sequencing of baculovirus,baculovirus gene function has become a hot topic in the field of baculovirus.Therefore,it is of great significance to study baculovirus gene function.The Autographa californica multiple,Nucleopolyhedroviruses is a representative species of baculovirus.At present,there are still many specific functions of some genes are not clear.In this paper,using the ac82 gene as the research object,using the Red homologous recombination system and Bac-to-Bac system construction of ac82 gene deletion and recovery of recombinant virus,to investigate the effect of ac82 gene deletion on the baculovirus,which reveals the gene function.The main findings include the following:Using Red system,the 64 bp gene of Ac MNPV ac82 gene in Bacmid was replaced by chloramphenicol resistance gene?Cm?,and the recombinant virus v Ac82 KO was successfully constructed with the deletion of ac82 gene in Bacmid.Using Bac-to-Bac system,successfully building GFP-expressing wild-type virus v Ac^PH-GFP,deletion-type recombinant virus v Ac^82KO-GFP and recovery type recombinant virus v Ac^82Rep-PH-GFP.Transfection and infection.The recombinant virus transfected cells have fluorescence,that this study successfully constructed the recombinant virus with green fluorescent protein gene gfp and polygonal body protein gene polh.Transfection experiments showed that the deletion of ac82 gene recombinant virus v Ac^82KO-PH-GFP Bacmid transfected cells and no diffusion of virus,that the gene deletion virus particles can not be normal to infect other cells through budding nearby cells,speculated that ac82 gene deletion affects the virus membrane process.