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The microRNA pathway activation in insect cell model upon Autographa californica multiple nucleopolyhedrovirus infection
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作者 QIAOJIN JIA YUEJUN FU 《BIOCELL》 SCIE 2023年第3期627-645,共19页
Background:The immune system of insects exerts fundamentally different antiviral mechanisms than mammals.MicroRNAs(miRNAs)play vital roles in developing insect antiviral immunity.MiRNAs expression profiles of insects ... Background:The immune system of insects exerts fundamentally different antiviral mechanisms than mammals.MicroRNAs(miRNAs)play vital roles in developing insect antiviral immunity.MiRNAs expression profiles of insects changed significantly during baculovirus infection.Methods:Differential expression profiles of miRNAs in Spodoptera frugiperda were monitored by next-generation sequencing(NGS)and RT-qPCR during Autographa californica multiple nucleopolyhedrovirus(AcMNPV)infection.The transcription levels of genes were detected by RT-qPCR.The 50%tissue culture infective dose(TCID_(50))endpoint dilution assay was used to determine the proliferation of progeny virus.Results:NGS revealed that 49 miRNAs were differentially expressed in Sf9 cells,and 10 of them were significantly up-or down-regulated.Though RT-qPCR analysis,we observed the similar trends for the expression patterns of significantly differentially expressed miRNAs from NGS.Moreover,the transcription levels of core genes,Exportin5,Dicer1,and Argonaute1,in miRNA biogenesis pathways were significantly increased after AcMNPV infection.For five selected miRNAs,miR-34-5p could regulate the proliferation of baculovirus progeny virus and energy metabolism.Conclusion:The miRNAs biogenesis pathway in Sf9 cells plays an important role and may be stimulated to resist AcMNPV infection.This work provides evidence for the molecular mechanism of baculovirus-insect interaction and offers novel ideas and directions for green pest control technology. 展开更多
关键词 MICRORNAS Spodoptera frugiperda Autographa californica multiple nucleopolyhedrovirus Host-virus interaction
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Application of Spodoptera litura Nucleopolyhedrovirus for Crop Pest Control
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作者 曾维爱 谭琳 +4 位作者 李小一 李宏光 谢鹏飞 黄艳宁 胡秋龙 《Agricultural Science & Technology》 CAS 2013年第9期1303-1306,共4页
The laboratory bioassay and field control efficacy of Spodoptera litura nucleopolyhedrovirus(Spli NPV) Chenzhou strain were preliminarily examined. The efficient artificial propagation method was to feed the host la... The laboratory bioassay and field control efficacy of Spodoptera litura nucleopolyhedrovirus(Spli NPV) Chenzhou strain were preliminarily examined. The efficient artificial propagation method was to feed the host larvae with virus suspension,and the average mortality of the insects was 65.0%. The death peak of the pests appeared 4-8 d after virus infection. The high temperature, high humidity and poor light could help the virus infection and propagation. Filed control efficacy of Chenzhou strain was 86.6% in laboratory, which was better than of another commercial strain. The corrected control efficacy of this strain was 88.4% the field, which was higher than that of avermectin pesticide significantly. It was detected that the occlusion body(OB) concentration of the initial virus' s stock solution was 1.03×1011OBs/ml,and it was a strong SpliNPV strain, as it showed an excellent efficacy to control the pest Spodoptera litura, and thus there will be a good prospect of application and development of this SpliNPV strain. 展开更多
关键词 Spodoptera litura nucleopolyhedrovirus Chenzhou strain BIOLOGY Control effect
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Encyclopedia of Autographa californica Nucleopolyhedrovirus Genes 被引量:2
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作者 David P. A. Cohen Martin Marek +2 位作者 Bryn G. Davies Just M. Vlak Monique M. van Oers 《Virologica Sinica》 SCIE CAS CSCD 2009年第5期359-414,共56页
The Autographa californica multiple capsid nucleopolyhedrovirus (AcMNPV) was the first baculovirus for which the complete nucleotide sequence became known. Since then 15 years lapsed and much research has been perform... The Autographa californica multiple capsid nucleopolyhedrovirus (AcMNPV) was the first baculovirus for which the complete nucleotide sequence became known. Since then 15 years lapsed and much research has been performed to elucidate putative functions of the annotated open reading frames of this virus and this endeavour is still ongoing. AcMNPV is the most well-known and well-studied baculovirus species, not in the least for its application as a vector for the high-level expression of foreign genes in insect cells. This article is the first monograph of a single baculovirus and gives a current overview of what is known about the 151 AcMNPV ORFs, including (putative) function and temporal and spatial presence of transcripts and protein. To date 60 ORFs have a proven function, another 19 ORFs have homologs for which functions are known in other baculoviruses and 72 ORFs are still enigmatic. This paper should assist the reader in quickly finding the essentials of AcMNPV. 展开更多
关键词 BACULOVIRUS Autographa californica multiple capsid nucleopolyhedrovirus(AcMNPV) Functional genomics Review
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The ORF 113 of Heliocoverpa armigera Single Nucleopolyhedrovirus Encodes a Functional Fibroblast Growth Factor 被引量:2
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作者 Xiang LI Chang-yong LIANG Jian-hua SONG Xin-wen CHEN 《Virologica Sinica》 SCIE CAS CSCD 2008年第5期321-329,共9页
Fibroblast growth factor (FGF) is a key regulator of developmental processes. A FGF homolog (vFGF) is found in all lepidopteran baculoviruses. Autographa californica nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (... Fibroblast growth factor (FGF) is a key regulator of developmental processes. A FGF homolog (vFGF) is found in all lepidopteran baculoviruses. Autographa californica nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV) vFGFs are chemotactic factors. Here we analyzed the vfgf of Helicoverpa armigera NPV (HearNPV), a group II NPV. The HearNPV vfgf transcripts were detected from 18 to 96 h post-infection (hpi) of Hz-AM1 cells with HearNPV and encoded a 36 kDa protein, which was secreted into the culture medium. HearNPV vFGF had strong affinity to heparin, a property important for FGF signaling via an FGF receptor. Unlike its AcMNPV homolog, HearNPV vFGF specially chemoattracted Hz-AM1, but not other insect cells such as Sf9 and Se-UCR and not the mammalian cells 293 and HepG2. HearNPV vFGF is also associated with the envelope of BV but is absent in occlusion-derived virus, which coordinated to the chemotatic activity analysis. 展开更多
关键词 Helicoverpa armigera nucleopolyhedrovirus (HearNPV) Fibroblast growth factor (FGF) BACULOVIRUS Open reading frame 113 (ORF 113)
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Putative Phosphorylation Sites On WCA Domain of HA2 Is Essential For Helicoverpa armigera Single Nucleopolyhedrovirus Replication
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作者 Yi-pin Lv Qian Wang +4 位作者 Chun-chen Wu Rong-juan Pei Yuan Zhou Yun Wang Xin-wen Chen 《Virologica Sinica》 SCIE CAS CSCD 2011年第4期245-251,共7页
Protein phosphorylation is one of the most common post-translational modification processes that play an essential role in regulating protein functionality.The Helicoverpa armigera single nucleopolyhedrovirus (HearNPV... Protein phosphorylation is one of the most common post-translational modification processes that play an essential role in regulating protein functionality.The Helicoverpa armigera single nucleopolyhedrovirus (HearNPV) orf2-encoded nucleocapsid protein HA2 participates in orchestration of virus-induced actin polymerization through its WCA domain,in which phosphorylation status are supposed to be critical in respect to actin polymerization.In the present study,two putative phosphorylation sites (232Thr and 250Ser) and a highly conserved Serine (245Ser) on the WCA domain of HA2 were mutated,and their phenotypes were characterized by reintroducing the mutated HA2 into the HearNPV genome.Viral infectivity assays demonstrated that only the recombinant HearNPV bearing HA2 mutation at 245Ser can produce infectious virions,both 232Thr and 250Ser mutations were lethal to the virus.However,actin polymerization assay demonstrated that all the three viruses bearing HA2 mutations were still capable of initiating actin polymerization in the host nucleus,which indicated the putative phosphorylation sites on HA2 may contribute to HearNPV replication through another unidentified pathway. 展开更多
关键词 Helicoverpa armigera single nucleopolyhedrovirus (HearNPV) Actin polymerization Protein phosphorylation N-WASP
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Entry into Midgut Epithelial Cells is a Key Step in the Selection of Genotypes in a Nucleopolyhedrovirus
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作者 Gabriel Clavijo Trevor Williams +2 位作者 Delia Muoz Miguel López-Ferber Primitivo Caballero 《Virologica Sinica》 SCIE CAS CSCD 2009年第4期350-358,共9页
An isolate of the Spodoptera frugiperda multiple nucleopolyhedrovirus comprises a stable proportion of deletion genotypes (e.g., SfNIC-C), that lack pifl and pif2 rendering them noninfectious per os, and that surviv... An isolate of the Spodoptera frugiperda multiple nucleopolyhedrovirus comprises a stable proportion of deletion genotypes (e.g., SfNIC-C), that lack pifl and pif2 rendering them noninfectious per os, and that survive by complementation with a complete genotype (SfNIC-B) in coinfected cells. To determine whether selection for particular ratios of complete and deletion genotypes occurs mainly during the establishment of the primary infection in insect midgut cells or during subsequent systemic infection, we examined genotype frequencies in insects that fed on OBs comprising different co-occluded mixtures of genotypes. Dramatic changes in genotype frequencies were observed between the OB inoculum and budded virus (BV) samples taken from larvae inoculated with OBs comprising 10% SfNIC-B + 90% SfNIC-C indicating that a marked reduction of SfNIC-C genotype had occurred in the insect midgut due to the immediate elimination of all OBs that originated from cells that had been infected only by SfNIC-C. In contrast, immediate changes were not observed in OBs comprising mixtures of 50% SfNIC-B + 50% SfNIC-C or those comprising 10% SfNIC-B + 90% SfNIC-C as most of the OBs in these mixtures originated from cells that had been infected by both genotypes. Subsequent changes in genotypic frequencies during five days of systemic infection were fairly small in magnitude for all genotypic mixtures. We conclude that the prevalence of defective genotypes in the SfNIC population is likely determined by a balance between host selection against OBs produced in cells infected by SfNIC-C alone and within-host selection for fast-replicating deletion genotypes. The strength of intra-host selection is likely modulated by changes in MOI during the infection period. 展开更多
关键词 nucleopolyhedrovirus Defective genotypes Infection SELECTION
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The piRNA pathway is required for nucleopolyhedrovirus replication in Lepidoptera
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作者 Junming Xia Shigang Fei +7 位作者 Hongyun Wu Yifan Yang Wensheng Yu Mengmeng Zhang Yiyao Guo Luc Swevers Jingchen Sun Min Feng 《Insect Science》 SCIE CAS CSCD 2023年第5期1378-1392,共15页
The Piwi-interacting RNA(piRNA)pathway has been shown to be involved in the antiviral defense against RNA viruses,especially in mosquitoes,but its universality has been questioned.Here,we used the Bombyx mori nucleopo... The Piwi-interacting RNA(piRNA)pathway has been shown to be involved in the antiviral defense against RNA viruses,especially in mosquitoes,but its universality has been questioned.Here,we used the Bombyx mori nucleopolyhedrovirus(BmNPV)-infected silkworm as a model to explore the effects of the key factors of piRNA pathway,BmAgo3 and Siwi,on replication of a large DNA virus(belonging to the family of Baculoviridae).We demonstrated that BmAgo3 and Siwi could promote the replication of BmNPV through both overexpression and knockdown experiments in BmN cell lines and silkworm larvae.In addition,we also studied the effect of PIWI-class genes on Autographa californica nucleopolyhedrovirus(AcMNPV)replication in the Spodoptera frugiperda cell line Sf9.By knocking down the expression of PIWI-class genes in Sf9,we found that Piwi-like-1 and Piwi-like-2-3 could inhibit AcMNPV replication,while Piwi-like-4-5 promoted virus replication.Our study provides compelling evidence that the piRNA pathway affects host infection by exogenous viruses in Lepidoptera.Also,our results reflect the diversity of the roles of PIWI-class genes in virus infection of the host across species.This study is the first to explore the interaction of PIWI-class proteins with DNA viruses,providing new insights into the functional roles of the piRNA pathway. 展开更多
关键词 BmAgo3 Bombyx mori nucleopolyhedrovirus Piwi-interacting RNA Siwi
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Expression and immunocytochemical analysis of Autographa californica nucleopolyhedrovirus (AcMNPV) orf74 gene 被引量:1
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作者 SHI-HENG AN ZHONG-JIANG GUO XIN-MING YIN 《Insect Science》 SCIE CAS CSCD 2006年第5期349-354,共6页
Autographa californica nucleopolyhedrovirus off74 (Ac74) is located between 62 311 and 63 108bp in the AcMNPV genome, which encodes 265 amino acid residues with a predicted 31 kDa molecular weight. The homologues of... Autographa californica nucleopolyhedrovirus off74 (Ac74) is located between 62 311 and 63 108bp in the AcMNPV genome, which encodes 265 amino acid residues with a predicted 31 kDa molecular weight. The homologues of Ac74 were searched using BLASTP in protein databases, GenBank/EMBL and SWISS-PROT. The result revealed that deduced Ac74 protein was homologous to the predicted products from 10 lepidoptera NPV ORFs. The multiple sequence alignments of Ac74 and its 10 homologues manifested only one amino acid residue was completely conserved. The transcript analysis revealed that the transcript of Ac74 was detected from 24-72 hours post-infection (hpi). The product of Ac74 was detected at 24 hpi and lasted until 72 hpi by Western blot using anti-Ac74 antiserum, consistent with reverse transcriptase polymerase chain reaction results. These results suggested Ac74 was expressed during the later stages of infection. The product of Ac74 was 31kDa in size, consistent with predicted molecular weight. The subcellular localization of Ac74 proteins manifested Ac74 protein in the cytoplasm, and was hardly present in the nucleus at 24 hpi. The fluorescence was also observed in polyhedra, except cytoplasm at 72 hpi. Together, Ac74 is a functional protein with 31kDa molecular weight and is located in the cytoplasm and the polyhedra. 展开更多
关键词 Ac74 Autographa californica EXPRESSION nucleopolyhedrovirus sequence analysis subcellular location transcript analysis
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Analysis of a late gene, orfl01 from Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus 被引量:1
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作者 SHI-HENG AN LI-PING XING +1 位作者 V. SHYAM KUMAR CHUAN-XI ZHANG 《Insect Science》 SCIE CAS CSCD 2005年第5期335-340,共6页
Helicoverpa armigera single nucleocapsid nucleopolyhedrovirns open reading frame 101 (ha101) is 762 nts in length and encodes a 254 amino acid peptide with predicted 29 kDa molecular weight. The homologues of ha101 ... Helicoverpa armigera single nucleocapsid nucleopolyhedrovirns open reading frame 101 (ha101) is 762 nts in length and encodes a 254 amino acid peptide with predicted 29 kDa molecular weight. The homologues of ha101 were explored using BLASTP searching tool in the updated GenBank/EMBL and SWlSS-PROT databases. The results showed that the homologues of ha101 were present in all the completely sequenced lepidopteran nucleopolyhedrovirnses and granulovirnses, suggesting that ha101 might be a functional gene associated with their lepidopteran hosts. Sequence alignment of ha101 and its homologues revealed that 10 amino acids were completely conserved. RT-PCR analysis of ha101 manifested that the transcript of ha101 was first detected at 24 hpi and remained detectable at up to 122 hpi, suggesting that ha101 was transcribed during late stages of infection. Ha101 was expressed using Bac to Bac system in Tn5B-1-4 cells. The product of ha101 expressed in Tn5B-1-4 cells was approximately 29kDa, consistent with the predicted molecular weight, and the results were confirmed by western blot analysis. The subcellular localization indicated that ha101 was aggregated along nuclear envelope during the early stages of infection and spread out to the entire nucleus including virogenic stroma in late stages of infection, suggesting that ha101 may play a specific role in virion assembly process or virogenic stroma arrangement. 展开更多
关键词 expression ha101 Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus sequence analysis RT-PCR subcellular localization
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Construction of a One-Vector Multiplex CRISPR/Cas9 Editing System to Inhibit Nucleopolyhedrovirus Replication in Silkworms 被引量:1
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作者 Zhanqi Dong Qi Qin +6 位作者 Zhigang Hu Peng Chen Liang Huang Xinling Zhang Ting Tian Cheng Lu Minhui Pan 《Virologica Sinica》 SCIE CAS CSCD 2019年第4期444-453,共10页
Recently the developed single guide(sg)RNA-guided clustered regularly interspaced short palindromic repeats/associated protein 9 nuclease(CRISPR/Cas9) technology has opened a new avenue for antiviral therapy. The CRIS... Recently the developed single guide(sg)RNA-guided clustered regularly interspaced short palindromic repeats/associated protein 9 nuclease(CRISPR/Cas9) technology has opened a new avenue for antiviral therapy. The CRISPR/Cas9 system uniquely allows targeting of multiple genome sites simultaneously. However, there are relatively few applications of CRISPR/Cas9 multigene editing to target insect viruses. To address the need for sustained delivery of a multiplex CRISPR/Cas9-based genome-editing vehicle against insect viruses, we developed a one-vector(pSL1180-Cas9-U6-sgRNA) system that expresses multiple sgRNA and Cas9 protein to excise Bombyx mori nucleopolyhedrovirus(BmNPV) in insect cells.We screened the immediate-early-1 gene(ie-1), the major envelope glycoprotein gene(gp64), and the late expression factor gene(lef-11), and identified multiple sgRNA editing sites through flow cytometry and viral DNA replication analysis. In addition, we constructed a multiplex editing vector(PSL1180-Cas9-sgIE1-sgLEF11-sgGP64, sgMultiple) to efficiently regulate multiplex gene-editing and inhibit BmNPV replication after viral infection. This is the first report of the application of a multiplex CRISPR/Cas9 system to inhibit insect virus replication. This multiplex system can significantly enhance the potential of CRISPR/Cas9-based multiplex genome engineering in insect virus. 展开更多
关键词 Bombyx mori Bombyx mori nucleopolyhedrovirus(BmNPV) ANTIVIRAL therapeutic CRISPR/Cas9 MULTIGENE editing
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Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus(BmNPV) 被引量:1
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作者 Xing-wei XIANG Rui YANG +4 位作者 Lin CHEN Xiao-long HU Shao-fang YU Cui-ping CAO Xiao-feng WU 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2012年第2期111-117,共7页
In the late phase of Bombyx mori nucleopolyhedrovirus(BmNPV)infection,a large amount of polyhedra appear in the infected cell nucleolus,these polyhedra being dense protein crystals protecting the incorporated virions ... In the late phase of Bombyx mori nucleopolyhedrovirus(BmNPV)infection,a large amount of polyhedra appear in the infected cell nucleolus,these polyhedra being dense protein crystals protecting the incorporated virions from the harsh environment.To investigate whether the foreign protein could be immobilized into the polyhedra of BmNPV,two recombinant baculoviruses were generated by a novel BmNPV polyhedrin-plus(polh +)Bac-to-Bac system,designated as vBmBac(polh +)-enhanced green fluorescent protein(EGFP)and vBmBac(polh +)-LacZ,which can express the polyhedrin and foreign protein simultaneously.Light microscopy analysis showed that all viruses produced polyhedra of normal appearance.Green fluorescence can be apparently detected on the surface of the vBmBac(polh +)-EGFP polyhedra,but not the BmNPV polyhedra.Fluorescence analysis and anti-desiccation testing confirmed that EGFP was embedded in the polyhedra.As expected,the vBmBac(polh +)-LacZ polyhedra contained an amount of LacZ and had a higherβ-galactosidase activity.Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)and Western blotting were also performed to verify if the foreign proteins were immobilized into polyhedra.This study provides a new inspiration for efficient preservation of useful proteins and development of new pesticides with toxic proteins. 展开更多
关键词 Bombyx mori nucleopolyhedrovirus POLYHEDRA Foreign protein Protein immobilization
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P33 of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus is a functional homolog of AcP33 被引量:1
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作者 Wenhua Kuang Huanyu Zhang +4 位作者 Dianhai Hou Manli Wang Fei Deng Hualin Wang Zhihong Hu 《Virologica Sinica》 SCIE CAS CSCD 2016年第4期346-349,共4页
Baculoviruses are insect-specific viruses with a circular double-stranded DNA genome ranging in size from 80-180 kb (Lu et al., 2012). Two distinct types of viri- ons have been identified during the infectious cycle... Baculoviruses are insect-specific viruses with a circular double-stranded DNA genome ranging in size from 80-180 kb (Lu et al., 2012). Two distinct types of viri- ons have been identified during the infectious cycle of baculoviruses, namely budded virions (BVs) and occlu- sion-derived virions (ODVs). BVs mediate infection from cell to cell, while ODVs initiate oral infection in the insect midgut (Braunagel and Summers, 2007). 展开更多
关键词 P33 of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus is a functional homolog of AcP33 Figure NPV
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Systematic Analysis of 42 Autographa Californica Multiple Nucleopolyhedrovirus Genes Identifies An Additional Six Genes Involved in the Production of Infectious Budded Virus
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作者 Tong Chen Xiaoyan Duan +6 位作者 Hengrui Hu Yu Shang Yangbo Hu Fei Deng Hualin Wang Manli Wang Zhihong Hu 《Virologica Sinica》 SCIE CAS CSCD 2021年第4期762-773,共12页
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
关键词 Autographa californica multiple nucleopolyhedrovirus(AcMNPV) BV production Essential genes Dispensable genes Important genes
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Autographa Californica Multiple Nucleopolyhedrovirus orf13 Is Required for Efficient Nuclear Egress of Nucleocapsids
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作者 Xingang Chen Xiaoqin Yang +3 位作者 Chengfeng Lei Fujun Qin Xiulian Sun Jia Hu 《Virologica Sinica》 SCIE CAS CSCD 2021年第5期968-980,共13页
Autographa californica multiple nucleopolyhedrovirus(Ac MNPV)orf13(ac13)is a conserved gene in all sequenced alphabaculoviruses.However,its function in the viral life cycle remains unknown.In this study,we found that ... Autographa californica multiple nucleopolyhedrovirus(Ac MNPV)orf13(ac13)is a conserved gene in all sequenced alphabaculoviruses.However,its function in the viral life cycle remains unknown.In this study,we found that ac13 was a late gene and that the encoded protein,bearing a putative nuclear localization signal motif,colocalized with the nuclear lamina.Deletion of ac13 did not affect viral genome replication,nucleocapsid assembly or occlusion body(OB)formation,but reduced virion budding from infected cells by approximately 400-fold compared with the wild-type virus.Deletion of ac13 substantially impaired the egress of nucleocapsids from the nucleus to the cytoplasm,while the OB morphogenesis was unaffected.Taken together,our results indicated that ac13 was required for efficient nuclear egress of nucleocapsids during virion budding,but was dispensable for OB formation. 展开更多
关键词 Autographa californica multiple nucleopolyhedrovirus(AcMNPV) orf13 Nucleocapsid egress OB morphogenesis
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Genome Characteristics of the Cyclophragma Undans Nucleopolyhedrovirus: A Distinct Species in Group Ⅰ of Alphabaculovirus
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作者 Zheng Zhu Jun Wang +11 位作者 Qianran Wang Feifei Yin Xiaoping Liu Dianhai Hou Lei Zhang Haizhou Liu Jiang Li Basil M. Arif Hualin Wang Fei Deng Zhihong Hu Manli Wang 《Virologica Sinica》 SCIE CAS CSCD 2018年第4期359-368,共10页
The Cyclophragma undans nucleopolyhedrovirus (CyunNPV), a potential pest control agent, was isolated from Cyclophragma undans (Lepidoptera: Lasiocampidae), an important forest pest. In the present study, we perfo... The Cyclophragma undans nucleopolyhedrovirus (CyunNPV), a potential pest control agent, was isolated from Cyclophragma undans (Lepidoptera: Lasiocampidae), an important forest pest. In the present study, we performed detailed genome analysis of CyunNPV and compared its genome to those of other Group I alphabaculoviruses. Sequencing of the CyunNPV genome using the Roche 454 sequencing system generated 142,900 bp with a G + C content of 45%. Genome analysis predicted a total of 147 hypothetical open reading frames comprising 38 baculoviral core genes, 24 lepidopteran baculovirus conserved genes, nine Group I Alphabaculovirus conserved genes, 71 common genes, and five genes that are unique to CyunNPV. In addition, the genome contains 13 homologous repeated sequences (hrs). Phylogenetic analysis groups CyunNPV under a distinct branch within clade "a" of Group I in the genus Alphabaculovirus. Unlike other members of Group I, CyunNPV harbors only nine of the 11 genes previously determined to be specific to Group I viruses. Furthermore, the CyunNPV lacks the tyrosine phosphatase gene and the ac30 gene. The CyunNPV F-like protein contains two insertions of continuous polar amino acids, one at the conventional fusion peptide and a second insertion at the pretransmembrane domain. The insertions are likely to affect the fusion function and suggest an evolutionary process that led to inactivation of the F-like protein. The above findings imply that CyunNPV is a distinct species under Group I Alphabaculovirus. 展开更多
关键词 Cyclophragma undans nucleopolyhedrovirus (CyunNPV) BACULOVIRUS Group I Alphabaculovirus F protein
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Characterization of the viral fibroblast growth factor homolog of Helicoverpa armigera single nucleopolyhedrovirus
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作者 Feifei Yin Ruikun Du +5 位作者 Wenhua Kuang Guang Yang Hualin Wang Fei Deng Zhihong Hu Manli Wang 《Virologica Sinica》 SCIE CAS CSCD 2016年第3期240-248,共9页
Fibroblast growth factor(FGF) is found throughout multicellular organisms; however, fgf homologs(vfgf) have only been identified among viruses in lepidopteran baculoviruses. The function of v FGFs from Group I alphaba... Fibroblast growth factor(FGF) is found throughout multicellular organisms; however, fgf homologs(vfgf) have only been identified among viruses in lepidopteran baculoviruses. The function of v FGFs from Group I alphabaculoviruses, including Autographa californica multiple nucleopolyhedrovirus(Ac MNPV) and Bombyx mori nucleopolyhedrovirus(Bm NPV), involves accelerated killing of infected larvae by both viruses. The v FGF of Group II alphabaculovirus is structurally different from that of Group I alphabaculovirus, with a larger C-terminal region and additional N-linked glycosylation sites. In this study, we characterized the Group II alphabaculovirus v FGF of Helicoverpa armigera single nucleopolyhedrovirus(Hear NPV). The transcription and expression of vfgf was detected at 3 h and 16 h post-infection in Hear NPV-infected cells. To further study v FGF function, we constructed vfgf-knockout and-repaired Hear NPV bacmids and investigated their affect in both cultured cells and insects. Deletion of vfgf had no effect on budded-virus production or viral DNA replication in cultured Hz AM1 cells. However, bioassays showed that Hear NPV vfgf deletion significantly increased the median lethal dose and delayed the median lethal time by ~12 h in the host insect when the virus was delivered orally. These results suggested that v FGF is an important virulent factor for HearN PV infection and propagation in vivo. 展开更多
关键词 deletion function Helicoverpa armigera single nucleopolyhedrovirus(HearNPV) infectivity vfgf
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Plutella xylostella granulovirus late gene promoter activity in the context of the Autographa californica multiple nucleopolyhedrovirus genome
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作者 He-Lin Ren Yuan Hu +1 位作者 Ya-Jun Guo Lu-Lin Li 《Virologica Sinica》 SCIE CAS CSCD 2016年第3期229-239,共11页
Within Baculoviridae, little is known about the molecular mechanisms of replication in betabaculoviruses, despite extensive studies in alphabaculoviruses. In this study, the promoters of nine late genes of the betabac... Within Baculoviridae, little is known about the molecular mechanisms of replication in betabaculoviruses, despite extensive studies in alphabaculoviruses. In this study, the promoters of nine late genes of the betabaculovirus Plutella xylostella granulovirus(Plxy GV) were cloned into a transient expression vector and the alphabaculovirus Autographa californica multiple nucleopolyhedrovirus(Ac MNPV) genome, and compared with homologous late gene promoters of Ac MNPV in Sf9 cells. In transient expression assays, all Plxy GV late promoters were activated in cells transfected with the individual reporter plasmids together with an Ac MNPV bacmid. In infected cells, reporter gene expression levels with the promoters of Plxy GV e18 and Ac MNPV vp39 and gp41 were significantly higher than those of the corresponding Ac MNPV or Plxy GV promoters,which had fewer late promoter motifs. Observed expression levels were lower for the Plxy GV p6.9,pk1, gran, p10 a, and p10 b promoters than for the corresponding Ac MNPV promoters, despite equal numbers of late promoter motifs, indicating that species-specific elements contained in some late promoters were favored by the native viral RNA polymerases for optimal transcription. The 8-nt sequence TAAATAAG encompassing the ATAAG motif was conserved in the Ac MNPV polh, p10,and pk1 promoters. The 5-nt sequence CAATT located 4 or 5 nt upstream of the T/ATAAG motif was conserved in the promoters of Plxy GV gran, p10 c, and pk1. The results of this study demonstrated that Plxy GV late gene promoters could be effectively activated by the RNA polymerase from Ac MNPV, implying that late gene expression systems are regulated by similar mechanisms in alphabaculoviruses and betabaculoviruses. 展开更多
关键词 Plutella xylostella granulovirus(PlxyGV) Autographa californica multiple nucleopolyhedrovirus(AcMNPV) late gene expression
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AcMNPV e18基因酵母双杂交诱饵载体构建和转录自激活检测 被引量:3
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作者 史瑞丽 周晓伟 黎路林 《湖北农业科学》 北大核心 2013年第10期2436-2438,2442,共4页
用PCR方法扩增苜蓿银纹夜蛾核多角体病毒(Autographa californica multiple nucleopolyhe-drovirus,AcMNPV)被膜蛋白ODV-E18基因,克隆至酵母双杂交诱饵载体pGBKT7构建诱饵质粒pG-BKT7-e18。将诱饵质粒分别转化酵母菌株Y187和AH109感受... 用PCR方法扩增苜蓿银纹夜蛾核多角体病毒(Autographa californica multiple nucleopolyhe-drovirus,AcMNPV)被膜蛋白ODV-E18基因,克隆至酵母双杂交诱饵载体pGBKT7构建诱饵质粒pG-BKT7-e18。将诱饵质粒分别转化酵母菌株Y187和AH109感受态细胞,被转化细胞在涂有X-gal的SD/-Trp营养缺陷型固体培养基上形成白色菌落;在SD/-Trp/-His和SD/-Trp/-Ade固体培养基上均不形成菌落,表明诱饵基因表达产物BD-E18在这两种细胞中都不能激活报告基因转录。pGBKT7-e18转化的Y187细胞在SD/-Trp营养缺陷型液体培养基中的生长速度与空载体转化细胞相同,显示BD-E18对酵母细胞无细胞毒性。结果表明,AcMNPV ODV-E18可能不直接参与对宿主细胞或病毒基因表达的调节,其编码基因可以作为诱饵基因通过筛查病毒宿主cDNA文库识别与其相互作用的蛋白质。 展开更多
关键词 苜蓿银纹夜蛾核多角体病毒(Autographa californica multiple nucleopolyhedrovirus AcMNPV) ODV-E18 酵母双杂交 自激活 细胞毒性
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茶刺蛾核型多角体病毒多角体的增殖动态与生产工艺 被引量:2
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作者 唐美君 殷坤山 +1 位作者 郭华伟 肖强 《植物保护》 CAS CSCD 北大核心 2015年第1期174-177,共4页
为了解茶刺蛾核型多角体病毒(Iragoides asciata nucleopolyhedrovirus IrfaNPV)的增殖特性、指导茶刺蛾病毒的生产,采用室内活体增殖法,测定了IrfaNPV多角体在茶刺蛾幼虫体内的增殖动态,并设计了工艺流程,进行了大量繁殖生产试验.结... 为了解茶刺蛾核型多角体病毒(Iragoides asciata nucleopolyhedrovirus IrfaNPV)的增殖特性、指导茶刺蛾病毒的生产,采用室内活体增殖法,测定了IrfaNPV多角体在茶刺蛾幼虫体内的增殖动态,并设计了工艺流程,进行了大量繁殖生产试验.结果显示,IrfaNPV多角体在虫体内的增殖动态符合Logistic曲线,饲毒后9~12 d为病毒快速增殖期.按设计的生产工艺进行病毒大量繁殖生产,虫尸中病毒多角体平均可达8.3×10^8 PIB/头,茧中病毒多角体含量平均为6.4×10^8 PIB/头.该工艺可用于今后茶刺蛾病毒的大量生产. 展开更多
关键词 茶刺蛾 核型多角体病毒 增殖动态 生产工艺
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Baculovirus per os Infectivity Factors Are Involved in HearNPV ODVs Infection of HzAM1 Cells in vitro 被引量:3
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作者 Ting JIANG Xiang LI +2 位作者 Jian-hua SONG Chang-yong LIANG Xin-wen CHEN 《Virologica Sinica》 SCIE CAS CSCD 2008年第1期25-30,共6页
Baculoviruses produce two viral phenotypes, the budded virus (BV) and the occlusion-derived virus (ODV). ODVs are released from occlusion bodies in the midgut where they initiate a primary infection. Due to the la... Baculoviruses produce two viral phenotypes, the budded virus (BV) and the occlusion-derived virus (ODV). ODVs are released from occlusion bodies in the midgut where they initiate a primary infection. Due to the lack of an in vitro system, the molecular mechanism of ODV infection is still unclear. Here we present data demonstrating that Helicoverpa armigera nucleopolyhedrovirus (HearNPV) ODV infected cultured Hz-AM1 cells in a pH dependent manner. The optimal pH for ODV infection was 8.5, which is same to that in the microvilli of midgut epithelial cells, the ODV native infection sites. Antibodies neutralization analysis indicated that four HearNPV oral infection essential genes p74, pif-l, pif-2 and pif-3 are also essential for HearNPV ODV infection in vitro. Thus, HearNPV-HzAM1 system can be used to analyze the mechanism of ODV entry. 展开更多
关键词 Helicoverpa armigera nucleopolyhedrovirus (HearNPV) Occlusion derived virus (ODV) per osinfectivity factor(pif) p74: Baculovirus
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