ABSTRACT
Bombyx mori nucleopolyhedrovirus (BmNPV) bm47 gene is found in all sequenced lepidopteran nucleopolyhedroviruses (NPVs). It is one of the core genes of NPVs. However, the role of bm47 in the biological cycle of NPV remains unknown. In this study, the Red recombination system was used to knock out bm47 from BmNPV to construct bm47-ko-Bacmid in E. coli BW25113 system. Then bm47 gene was introduced back to the viral genome using the Bac-to-Bac system to create the repair virus bm47-re-Bacmid. TCID50 assay and real-time PCR (qPCR) were used to evaluate the effects of bm47 deletion on viral DNA replication, gene transcription, and protein expression. qPCR results showed that bm47 knock-out had no significant effect on viral DNA replication. However, the qPCR results showed that bm47-ko-Bacmid significantly decreased the transcription levels of early gene lef-3, late gene vp39, and very late gene p10 at 48 h and 72 h after viral transfection of BmN cells (P < 0.05). This work will provide a foundation for further studies on the biological function of BmNPV bm47 in viral replication and transcription.
Subject(s)
Animals , Bombyx , Virology , Gene Deletion , Gene Expression Regulation, Viral , Nucleopolyhedroviruses , Genetics , Physiology , Transcription, Genetic , Viral Proteins , Genetics , Metabolism , Virus ReplicationABSTRACT
At present, microbe surface display system mainly involves phage surface display system, bacte- rial surface display system, yeast surface display system and virus surface display system. Baculovirus sur- face display system is a new type of eukaryote surface display system which developed based on deeply un- derstanding of construction and function of virus genome in recent years. Through fused expression with vi- ral capsid or membrane proteins exogenous peptides can be displayed on the surface of the virus and formed hedgehog-shape "fake virus". Baculovirus surface display system was characterized by safeness and high performance, furthermore, this system can complete post-translation processing and modification of protein to enhance the bioactivity of exogenous product. Combined with the author’s experimental work, this paper briefly introduces the mechanism and traits of this system and summarizes the newest research development on its application in the field of monoclonal antibody preparation, new-type vaccine development, genes transduction and genes therapy. It is believed that the system above may show extensive application through further improvement and optimization.