Study On Quantum Dots Labeling Of BmNPV BV And Analysis GP64 Signal Peptide

Bombyx mori nucleopolyhedrovirus(BmNPV)is the most serious pathogenic microorganism in the sericulture industry.It is a representative member of genus nuclear polyhedrosis virus of baculoviridae.Therefore,there are more baculoviruses currently studied.At present,there have been a lot of researches on the infection mechanism of BmNPV,but there are still many problems that need to be further explored for the specific invasion mechanism of BV.This paper mainly focuses on quantum dot-labeled BmNPV BV and studies GP64 signal peptide differences.Further studies revealed that the N-ter minal signal peptide of the BmNPV GP64 mediates the transport and secretion of proteins,and the C-ter minal transmembrane domain is anchored to the cell membrane.This study will provide a basic reference for the later experimental study of specific pathways for BmNPV BV entry and exit into host cells.The first chapter introduction: Introduced the baculovirus classification,infection mechanism,expression system,applications;the advantages and applications of quantum dots;introduced the baculovirus BmNPV and AcMNPV invasion host cells research status at home and abroad.Chapter two construction of quantum dot-tagged BmNPV BV: We first constructed a biotinylated recombinant baculovirus,followed by quantum dot labeling and laser scanning confocal laser microscopy to analyze the process of viral infection of host cells.In the process of quantum dot marking we have a new discovery that the AcMNPV quantum dot labeling method cannot be applied in BmNPV.Chapter three difference analysis of different GP64 signal peptides recombinant baculovirus: Our previous study found that signal peptide did not cleave during the post-processing of BmNPV GP64.The preli minary analysis may be due to the fact that BmNPV GP64 in the late stage is different from the Golgi-endoplasmic reticulum pathway biotin synthesis and secretion.Therefore,we analyzed the characteristics of the BmNPV GP64 signal peptide sequence and found that the recombinant viruses with different lengths of GP64 signal peptide have very different infectivity.Immunofluorescence analysis of the complete signal peptide GP64 is located on the cytoplasmic membrane.Chapter IV different lengths of GP64 signal peptides to guide eGFP localization and secretion analysis: based on the results of the previous chapter,the difference in the baculovirus characteristics of different GP64 signal peptides.We used the signal peptide+eGFP+TMD to model the membrane protein GP64,through transient transfection,analyzed the localization of eGFP in the cell by different signal peptides.The differences in the localization and secretion pathways will better explain why the AcMNPV and BmNPV marker methods differ and provide a basis for the specific pathway of BmNPV virion into and out of host cells.Through the above studies,we successfully constructed a variety of labeled viruses,and for the first time revealed that BmNPV GP64 may be different from the traditional secretion pathway,which provides a new reference for the further study of BmNPV access to host cells.