Investigating baculovirus’s host range through co-infect different baculoviruses

• Main Authors: YAN-CHANG ZHU, 朱彥昌
• Other Authors: Tzong Yuan Wu
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/53762331653070838744

碩士 === 中原大學 === 生物科技研究所 === 99 === Baculovirus is named after the host that it infects. Most of baculoviruses have their own specific host range, but some have wide host range that can infect many different insectspecies, like Auotographa californica multiple nucleopolyhedrovirus (AcMNPV) can infect 33 kind of insects and different kind of insect cell lines. Moreover, Maruca vitrata multiple nucleopolyhedrovirus (MaviMNPV) and Bombyx mori multiple nucleopolyhedrovirus (BmNPV) are two baculovirus that are similar in genome and structure. Despite the similarity in genome and structure, they can only infect their host range for instance AcMNPV infects sf21 cell line, BmNPV infects BmN cell line, and MaviMNPV infects NTU-MV532 cell line. We tried to find out the relationship between viruses and host their interaction. Different baculoviruses was incorporated with a different fluorescent reporter gene to co-infect Sf21,BmN and NTU-MV532 cell lines. Recombinant virus of AcMNPV and MaviMNPV was produced by cross infecting in Sf21 and NTU-MV532 which could infect both Sf21 and NTU-MV532 cell lines. Late gene expression of respective gene driven by polyhedrin promoter was studied by observing their fluorescent expression after single or co-infection with AcMNPV and MaviMNPV in Sf21 and NTU-MV532 cells lines at different time intervals. Co-infection of AcMNPV and MaviMNPV in Sf21 cell suggests that AcMNPV and MaviMNPV help each other’s gene expression in late phase, but blocks gene expression in NTU-MV532 cell line. Similar, result was observed in BmN and Sf21 cells which was co-infected with AcMNPV and BmNPV. Additionally, to study the influence of secretion factor in host range ,condition medium was prepared byultracentrifugation of medium infected with AcMNPV which was used to culture theMaviMNPV infected Sf21 cells.The result suggests that the secretion factor didn't play any role in influencing host range.QRT-PCR was performed to identify the genes that are essential for specific host range and several early and core genes were targeted for the study and their gene expression was measured in different time intervals.The expression of IE-1, IE-2 and lef-2 geneswere higher in co-infectedSf21cells with AcMNPV and MaviMNPV at 24 and 36 hpi; than single infection of AcMNPV. However, the expression of IE-1, IE-2 and lef-2 geneswas higher in co-infected Sf21 cells with AcMNPV and MaviMNPV than single infection of MaviMNPV. No gene expression of IE-1, IE-2 and lef-2 genes observed inNTU-MV532 cells co-infected with AcMNPV and MaviMNPV or AcMNPV alone but high expression of genes observed in NTU-MV532 cells infected with MaviMNPV alone.