The fall armyworm,Spodoptera frugiperda,which destroys many economic crops such as rice and maize,has recently invaded China.Insect viruses as biological control agents play important roles in killing pests.One potent...The fall armyworm,Spodoptera frugiperda,which destroys many economic crops such as rice and maize,has recently invaded China.Insect viruses as biological control agents play important roles in killing pests.One potential viral insecticide is the environmentally highly infective and virulent densovirus.We successfully rescued Junonia coenia densovirus(JcDV)using its infectious clone in different insect cell lines and larvae of three insect species.Results showed that the lysate of cultured insect cells transfected by the JcDV infectious clone killed the 2 nd instar S.frugiperda.The LD50 of homogenate from JcDV-infected Spodoptera litura to the 2 nd instar S.frugiperda(1.76×10^(8)viral genome copies per larva during 10 d post infection)was higher than that of the 2 nd instar S.litura(7.39×10^(7)Jc DV genome copies)or Helicoverpa armigera larvae(9.71×10^(7)JcDV genome copies).The LT50 of the S.litura homogenate(2.60×10^(9)viral genome copies each larva)to the 2 nd instar S.frugiperda was 6.96 d,longer than that of the S.litura(6.18 d)or the 2 nd instar H.armigera(5.94 d).JcDV could infect the fat body of H.armigera,but not S.frugiperda or S.litura.Although JcDV can infect all three lepidopteran species,their susceptibility to the virus differs.JcDV has great potential as a biological control agent against pests such as S.frugiperda.展开更多
The gene of the non-structure protein 2 （NS2） was cloned by PCR from the genome ofBombyx mori densovirus Zhenjiang strain （BmDNV-Z）, inserted into prokaryotic expression vector pET28a to construct recombinant plas...The gene of the non-structure protein 2 （NS2） was cloned by PCR from the genome ofBombyx mori densovirus Zhenjiang strain （BmDNV-Z）, inserted into prokaryotic expression vector pET28a to construct recombinant plasmid pET28a-NS2 and then expressed in bacteria Escherichia coli BL21 （DE3）. The expressed recombinant protein was identified by SDS-PAGE and Western blot analysis. Then, the recombinant protein was purified by Ni-NTA column, renatured and tested for enzyme activities. The purified NS2 protein exhibited a helicase activity unwinding double-stranded DNA substrates into single-strand primers, and higher unwinding activity to polarity substrate. Similarly, the purified NS2 protein possessed an ATPase activity and its enzyme activity was 0.276 μmol gg^-1 h^-1 in this study. The results indicated that the non- structure protein which encoded by the gene of BmDNV-Z NS2 possesses the biological activities of helicase and ATPase, and the helicase prefers to polarity substrates. Based on these results, it is speculated that the gene of BmDNV-Z NS2 plays an important role in the viral DNA replication.展开更多
为了探明浓核病毒镇江株(BmDNV-ZJ)感受性家蚕品种中肠组织不同部位对浓核病毒感受性差异的蛋白质组学机制,阐明家蚕浓核病毒的致病机理,应用等电聚焦(IFE),SDS-PAGE分离,2D P latinum软件以及质谱技术,对感受性家蚕品种中肠前、中、后...为了探明浓核病毒镇江株(BmDNV-ZJ)感受性家蚕品种中肠组织不同部位对浓核病毒感受性差异的蛋白质组学机制,阐明家蚕浓核病毒的致病机理,应用等电聚焦(IFE),SDS-PAGE分离,2D P latinum软件以及质谱技术,对感受性家蚕品种中肠前、中、后3个区段的组织蛋白进行了比较分析.结果表明:中肠前、中、后3个区段组织蛋白的点数、位置、点形等差异较小.在中肠前部和后部组织中找出的4个差异蛋白点,分别为脂蛋白前体、囊膜氢离子泵ATP合酶B亚基、α胰蛋白酶A链和肌动蛋白;在中肠中部找出的一个差异蛋白点为Kun itz型胰蛋白酶抑制剂A链.这些蛋白点可能在BmDNV-ZJ侵染感受性家蚕中肠组织细胞中起一定的作用.展开更多
A luciferase gene has been inserted into the recombinant plasmid PfDNV-pUC119 which contained partly deletion of genome of Periplanete fuliginosa densovirus (PfDNV). The recombinant plasmid with luciferase gene was co...A luciferase gene has been inserted into the recombinant plasmid PfDNV-pUC119 which contained partly deletion of genome of Periplanete fuliginosa densovirus (PfDNV). The recombinant plasmid with luciferase gene was co-transfected with pfDNV-pUC119 into Periplanele fuliginosa larvae and had a high luciferase gene expression in enteron of the transfected larvae.展开更多
基金supported by the National Key R&D Program of China(2017YFD0200400)the Natural Science Foundation of Hubei Province,China(2017CFB241)。
文摘The fall armyworm,Spodoptera frugiperda,which destroys many economic crops such as rice and maize,has recently invaded China.Insect viruses as biological control agents play important roles in killing pests.One potential viral insecticide is the environmentally highly infective and virulent densovirus.We successfully rescued Junonia coenia densovirus(JcDV)using its infectious clone in different insect cell lines and larvae of three insect species.Results showed that the lysate of cultured insect cells transfected by the JcDV infectious clone killed the 2 nd instar S.frugiperda.The LD50 of homogenate from JcDV-infected Spodoptera litura to the 2 nd instar S.frugiperda(1.76×10^(8)viral genome copies per larva during 10 d post infection)was higher than that of the 2 nd instar S.litura(7.39×10^(7)Jc DV genome copies)or Helicoverpa armigera larvae(9.71×10^(7)JcDV genome copies).The LT50 of the S.litura homogenate(2.60×10^(9)viral genome copies each larva)to the 2 nd instar S.frugiperda was 6.96 d,longer than that of the S.litura(6.18 d)or the 2 nd instar H.armigera(5.94 d).JcDV could infect the fat body of H.armigera,but not S.frugiperda or S.litura.Although JcDV can infect all three lepidopteran species,their susceptibility to the virus differs.JcDV has great potential as a biological control agent against pests such as S.frugiperda.
基金supported by the National Basic Research Program of China (2005CB121004)
文摘The gene of the non-structure protein 2 （NS2） was cloned by PCR from the genome ofBombyx mori densovirus Zhenjiang strain （BmDNV-Z）, inserted into prokaryotic expression vector pET28a to construct recombinant plasmid pET28a-NS2 and then expressed in bacteria Escherichia coli BL21 （DE3）. The expressed recombinant protein was identified by SDS-PAGE and Western blot analysis. Then, the recombinant protein was purified by Ni-NTA column, renatured and tested for enzyme activities. The purified NS2 protein exhibited a helicase activity unwinding double-stranded DNA substrates into single-strand primers, and higher unwinding activity to polarity substrate. Similarly, the purified NS2 protein possessed an ATPase activity and its enzyme activity was 0.276 μmol gg^-1 h^-1 in this study. The results indicated that the non- structure protein which encoded by the gene of BmDNV-Z NS2 possesses the biological activities of helicase and ATPase, and the helicase prefers to polarity substrates. Based on these results, it is speculated that the gene of BmDNV-Z NS2 plays an important role in the viral DNA replication.
文摘为了探明浓核病毒镇江株(BmDNV-ZJ)感受性家蚕品种中肠组织不同部位对浓核病毒感受性差异的蛋白质组学机制,阐明家蚕浓核病毒的致病机理,应用等电聚焦(IFE),SDS-PAGE分离,2D P latinum软件以及质谱技术,对感受性家蚕品种中肠前、中、后3个区段的组织蛋白进行了比较分析.结果表明:中肠前、中、后3个区段组织蛋白的点数、位置、点形等差异较小.在中肠前部和后部组织中找出的4个差异蛋白点,分别为脂蛋白前体、囊膜氢离子泵ATP合酶B亚基、α胰蛋白酶A链和肌动蛋白;在中肠中部找出的一个差异蛋白点为Kun itz型胰蛋白酶抑制剂A链.这些蛋白点可能在BmDNV-ZJ侵染感受性家蚕中肠组织细胞中起一定的作用.
文摘A luciferase gene has been inserted into the recombinant plasmid PfDNV-pUC119 which contained partly deletion of genome of Periplanete fuliginosa densovirus (PfDNV). The recombinant plasmid with luciferase gene was co-transfected with pfDNV-pUC119 into Periplanele fuliginosa larvae and had a high luciferase gene expression in enteron of the transfected larvae.
