Baculoviruses have been widely used as a vector for expressing foreign genes.Among numerous baculoviruses,Autographa californica multiple nucleopolyhedrovirus(AcMNPV)is the most frequently used and it encodes 155 open...Baculoviruses have been widely used as a vector for expressing foreign genes.Among numerous baculoviruses,Autographa californica multiple nucleopolyhedrovirus(AcMNPV)is the most frequently used and it encodes 155 open reading frames(ORFs).Here,we systematically investigated the impact of 42 genes of AcMNPV on the production of infectious budded viruses(BVs)by constructing gene-knockout bacmids and subsequently conducting transfection and infection assays.The results showed that among the 39 functionally unverified genes and 3 recently reported genes,36 are dispensable for infectious BV production,as the one-step growth curves of the gene-knockout viruses were not significantly different from those of the parental virus.Three genes(ac62,ac82 and ac106/107)are essential for infectious BV production,as deletions thereof resulted in complete loss of infectivity while the repaired viruses showed no significant difference in comparison to the parental virus.In addition,three genes(ac13,ac51 and ac120)are important but not essential for infectious BV production,as gene-knockout viruses produced significantly lower BV levels than that of the parental virus or repaired viruses.We then grouped the 155 AcMNPV genes into three categories(Dispensable,Essential,or Important for infectious BV production).Based on our results and previous publications,we constructed a schematic diagram of a potential mini-genome of AcMNPV,which contains only essential and important genes.The results shed light on our understanding of functional genomics of baculoviruses and provide fundamental information for future engineering of baculovirus expression system.展开更多
基金supported by the National Natural Science Foundation of China(21890753,21988101 to Dehui Deng,91853101 to Fangjun Wang,and 91845106 to Liang Yu)the Strategic PriorityResearch Program of the Chinese Academy of Sciences(XDB36030200 to Dehui Deng)the Youth Innovation Promotion Association of the Chinese Academy of Sciences(Y201936 to Dehui Deng,Y201750 to Yangbo Hu)。
基金This research was supported by the grants from the National Natural Science Foundation of China(No.31872640)the Key Research Program of Frontier Sciences of the Chinese Academy of Sciences(grant no.QYZDJ-SSW-SMC021)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB11030400).
文摘Baculoviruses have been widely used as a vector for expressing foreign genes.Among numerous baculoviruses,Autographa californica multiple nucleopolyhedrovirus(AcMNPV)is the most frequently used and it encodes 155 open reading frames(ORFs).Here,we systematically investigated the impact of 42 genes of AcMNPV on the production of infectious budded viruses(BVs)by constructing gene-knockout bacmids and subsequently conducting transfection and infection assays.The results showed that among the 39 functionally unverified genes and 3 recently reported genes,36 are dispensable for infectious BV production,as the one-step growth curves of the gene-knockout viruses were not significantly different from those of the parental virus.Three genes(ac62,ac82 and ac106/107)are essential for infectious BV production,as deletions thereof resulted in complete loss of infectivity while the repaired viruses showed no significant difference in comparison to the parental virus.In addition,three genes(ac13,ac51 and ac120)are important but not essential for infectious BV production,as gene-knockout viruses produced significantly lower BV levels than that of the parental virus or repaired viruses.We then grouped the 155 AcMNPV genes into three categories(Dispensable,Essential,or Important for infectious BV production).Based on our results and previous publications,we constructed a schematic diagram of a potential mini-genome of AcMNPV,which contains only essential and important genes.The results shed light on our understanding of functional genomics of baculoviruses and provide fundamental information for future engineering of baculovirus expression system.
