The polyhedrin (polh) gene is often used to analyse evolution of baculovirus. In this report, the polh of Antheraea pernyi nucleopolyhedro-virus (AnpeNPV) was cloned and sequenced. The Open reading frame (ORF) of the ...The polyhedrin (polh) gene is often used to analyse evolution of baculovirus. In this report, the polh of Antheraea pernyi nucleopolyhedro-virus (AnpeNPV) was cloned and sequenced. The Open reading frame (ORF) of the AnpeNPV consists of 738 nucleotides encoding 245 amino acids with molecular masses of 29 kDa. The deduced amino acids were significant homol-ogy with other baculoviruses, such as Attacus ricini NPV (ArNPV) and Autographa californica NPV (AcNPV). A strongly hydrophilic region was predicted at positions from 30 to 50 of the An-peNPV Polh protein by bioinformatics analysis. Expression of the polh gene of AnpeNPV in E. coli was examined by SDS–PAGE, Western blot and Mass-spectrum analysis. The result showed that the bacterium expression system was suitable for the virus gene expression. It indi-cated that the products of the polh gene ex-pressed in this system can be easier to use for raising antibodies.展开更多
家蚕核型多角体病毒(Bombyx mori nucleopolyhe⁃drovirus,BmNPV)是家蚕的主要病原之一,由BmNPV引发的血液型脓病对蚕桑产业的危害极大。对BmNPV3个地理分离株BmNPV⁃Zhenjiang、BmNPV⁃Baoshan和BmNPV⁃Sanya进行全基因组测序分析。在3个...家蚕核型多角体病毒(Bombyx mori nucleopolyhe⁃drovirus,BmNPV)是家蚕的主要病原之一,由BmNPV引发的血液型脓病对蚕桑产业的危害极大。对BmNPV3个地理分离株BmNPV⁃Zhenjiang、BmNPV⁃Baoshan和BmNPV⁃Sanya进行全基因组测序分析。在3个地理株基因组序列之间检测到位于编码区的90个单核苷酸多态性(single⁃nucleotide pol⁃ymorphism,SNP)位点和7个插入⁃缺失多态性(insertion⁃de⁃letion,InDel)位点,导致26个杆状病毒核心基因中的21个所编码的氨基酸序列发生了突变。这21个基因所涉及的功能包括:杆状病毒基因组的复制与转录、病毒粒子的组装以及病毒的侵染。此外,检测了3个地理株对菁松×皓月2龄幼蚕的致死中浓度(LC_(50)),并分析了ie⁃1、polh和p143这3个基因在菁松×皓月5龄幼蚕中肠组织中的相对表达水平。BmNPV⁃Zhenjiang与BmNPV⁃Baoshan的LC_(50)无显著差异,而BmNPV⁃Sanya的LC_(50)显著高于BmNPV⁃Zhenjiang和BmNPV⁃Baoshan(P≤0.01)。在幼蚕感染3个地理株48 h、72 h和96 h后,BmNPV⁃Zhenjiang与BmNPV⁃Baoshan中ie⁃1、polh和p143的相对表达水平无显著差异,而BmNPV⁃Sanya中polh、ie⁃1和p143的相对表达水平显著低于BmNPV⁃Zhenjiang和BmNPV⁃Baoshan(P≤0.01)。分析结果表明,BmNPV⁃Zhenjiang与BmNPV⁃Baoshan的毒力不具有较大差异,而BmNPV⁃Sanya的毒力弱于BmNPV⁃Zhenjiang和BmNPV⁃Baoshan。3个地理株之间核心基因的差异可能与其毒力差异存在一定关系,研究结果为阐明造成BmNPV不同地理分离株产生毒力差异的分子机制提供了初步的线索。展开更多
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
The combinations of Hyphantria cunea NPV(HcNPV) and three different fluorescent brighteners BA,VBL,CBS-X were added to the artificial diet and the influence of synergists in concentration 25 ℃ and 14L/10D(h).The resu...The combinations of Hyphantria cunea NPV(HcNPV) and three different fluorescent brighteners BA,VBL,CBS-X were added to the artificial diet and the influence of synergists in concentration 25 ℃ and 14L/10D(h).The results showed that the fluorescent brighteners BA and VBL had higher insecticidal activity,CBS-X had not significant synergistic effect.展开更多
文摘The polyhedrin (polh) gene is often used to analyse evolution of baculovirus. In this report, the polh of Antheraea pernyi nucleopolyhedro-virus (AnpeNPV) was cloned and sequenced. The Open reading frame (ORF) of the AnpeNPV consists of 738 nucleotides encoding 245 amino acids with molecular masses of 29 kDa. The deduced amino acids were significant homol-ogy with other baculoviruses, such as Attacus ricini NPV (ArNPV) and Autographa californica NPV (AcNPV). A strongly hydrophilic region was predicted at positions from 30 to 50 of the An-peNPV Polh protein by bioinformatics analysis. Expression of the polh gene of AnpeNPV in E. coli was examined by SDS–PAGE, Western blot and Mass-spectrum analysis. The result showed that the bacterium expression system was suitable for the virus gene expression. It indi-cated that the products of the polh gene ex-pressed in this system can be easier to use for raising antibodies.
基金funded by the Natural Science Foundation of Shanxi Province,Grant No.201801D121193the Central Government Guiding Local Science and Technology Development Fund Project,Grant No.YDZJSX2022A001.
文摘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.
文摘The combinations of Hyphantria cunea NPV(HcNPV) and three different fluorescent brighteners BA,VBL,CBS-X were added to the artificial diet and the influence of synergists in concentration 25 ℃ and 14L/10D(h).The results showed that the fluorescent brighteners BA and VBL had higher insecticidal activity,CBS-X had not significant synergistic effect.
