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Genome sequence of a Papaya ringspot virus from khejri (Prosopis cineraria) transcriptome from India 来自印度番木瓜环斑病毒转录组的基因组序列
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_409
M. Prajapati, Jitender Singh, Pankaj Kumar, V. Baranwal
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引用次数: 0
Natural infection of cape gooseberry (Physalis peruviana) by the potyvirus Tamarillo leaf malformation virus (TaLMV). 柽柳叶畸形病毒(TaLMV)对猕猴桃自然侵染的研究。
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_211
Erika Corrales-Cabra, Andrea Restrepo Escobar, Mónica Higuita Valencia, Yuliana Gallo García, Mauricio Marín Montoya, Pablo Gutiérrez Sánchez

Tamarillo leaf malformation virus (TaLMV) is a potyvirus first discovered in cape gooseberry fields in Eastern, and South-western Antioquia. This virus is responsible for a very damaging disease that has resulted in significant reductions in yields and cultivated area for this crop in Colombia. Tamarillo is frequently co-cultivated with other solanaceous plants but no evidence for cross-pathogenicity of TaLMV has been found until now. In this work, we report a natural infection of cape gooseberry (Physalis peruviana L.) by TaLMV. Infection by TaLMV was detected by RNAseq screening of cape gooseberry fields and confirmed by RT-qPCR and Sanger sequencing. The sequenced genome is 99.3% identical to previously sequenced TaLMV isolates, and evidence suggests that it can accumulate at high loads in this new reported host. RT-qPCR analysis indicates that TaLMV is already widely distributed, can naturally infect other solanaceous hosts and may become an emerging threat to the cape gooseberry agroindustry, the second most important exotic fruit export in Colombia. Keywords: high-throughput sequencing; plant virology; Potyviridae; RT-qPCR; Solanaceae.

柽柳叶畸形病毒(Tamarillo leaf malformation virus, TaLMV)是一种首次在安蒂奥基亚省东部和西南部的醋栗田发现的痘病毒。这种病毒造成了一种极具破坏性的疾病,导致哥伦比亚这种作物的产量和种植面积大幅减少。柽柳经常与其他茄类植物共栽培,但迄今尚未发现TaLMV的交叉致病性证据。在这项工作中,我们报告了一种由TaLMV自然侵染的醋栗(Physalis peruviana L.)。采用RNAseq筛选法对猕猴桃地进行了TaLMV感染检测,并通过RT-qPCR和Sanger测序进行了验证。测序的基因组与先前测序的TaLMV分离株有99.3%的相同,并且有证据表明它可以在这个新报道的宿主中大量积累。RT-qPCR分析表明,TaLMV已经广泛分布,可以自然感染其他茄类寄主,并可能成为哥伦比亚第二大外来水果出口的开普醋莓农工业的新威胁。关键词:高通量测序;植物病毒学;Potyviridae;RT-qPCR;茄科。
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引用次数: 1
Selective human cells in vitro transduction with porcine endogenous retrovirus (PERV). 猪内源性逆转录病毒(PERV)体外转染人细胞的选择性研究。
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_201
Sabina Gałka, Ewa Nowak, Ilona Bednarek

Porcine xenograft transplantation raises concerns in humans about the risk of infection with porcine endogenous retroviruses (PERV) as they are an integral part of the pig genome and are therefore very difficult to exclude. In this study, for the first time, a relationship between the provirus genes sequences and released virions from pig cell line and the embedded sequence of this retrovirus in infected human cells was analyzed. PERV infection of human cells HEK-293 and HeLa and detection of PERV in pig PK-15 cells and supernatant were assessed by QPCR or RT-QPCR using primers specific for envA, envB, gag, pol genes and LTR region. Sequence analysis was performed at the DNA level and changes in the amino acid sequence were deduced in silico. Fifty nucleotide substitutions (45 in pol, 3 in gag and one each in envA and envB) were detected and most of these were heterozygous (42), which were present mainly in PK-15 cells. Our results show that sequence of the pol gene and the Pol protein is less conserved compared to the other PERV genes and PERV with some polymorphisms were not released from pig cells or/and do not infect human cells. PERV virions with a homozygous allele system were released from PK-15 cells, although their sequence replicated on the basis of the heterozygous PERV provirus sequence in PK-15. The newly discovered selective transduction of human cells with PERV will be helpful in studying the characteristics and genetic variability of the retrovirus genes to ensure safe xenotransplantation. Keywords: PERV; porcine endogenous retroviruses; infection; genetic polymorphism; xenotransplantation.

猪异种移植物移植引起了人类对猪内源性逆转录病毒(PERV)感染风险的担忧,因为它们是猪基因组的一个组成部分,因此很难排除。本研究首次分析了猪细胞系中原病毒基因序列和释放的病毒粒子与被感染人细胞中该逆转录病毒的嵌入序列之间的关系。采用envA、envB、gag、pol基因和LTR区特异性引物,采用QPCR或RT-QPCR检测PERV对人HEK-293和HeLa细胞的感染情况,以及猪PK-15细胞和上清的PERV检测情况。在DNA水平上进行序列分析,并通过计算机推导出氨基酸序列的变化。共检测到50个核苷酸替换(pol 45个,gag 3个,envA和envB各1个),其中大部分是杂合的(42个),主要存在于PK-15细胞中。结果表明,与其他PERV基因相比,pol基因和pol蛋白的序列保守性较低,具有部分多态性的PERV不会从猪细胞中释放或感染人类细胞。具有纯合子等位基因系统的PERV病毒粒子从PK-15细胞中释放出来,尽管它们的序列是在PK-15的杂合子PERV原病毒序列的基础上复制的。新发现的PERV对人类细胞的选择性转导,将有助于研究逆转录病毒基因的特性和遗传变异性,确保异种移植的安全性。关键词:恋物癖;猪内源性逆转录病毒;感染;基因多态性;异种移植。
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引用次数: 0
First record of mosquito-borne Sindbis virus (genotype I) in the Czech Republic. 捷克共和国首次记录蚊媒信德比斯病毒(基因型I)。
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_313
Silvie Šikutová, Jan Mendel, Jakub Vojtíšek, Zdeněk Hubálek, Ivo Rudolf
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引用次数: 0
RT-qPCR based determination of SARS-CoV-2 in health care workers, Cologne, Germany – A pre-delta/pre-omicron proof of principle field study 基于RT-qPCR的SARS-CoV-2在德国科隆卫生保健工作者中的检测——一项delta前/组粒前的原理现场研究证明
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_403
Gina Spölgen, J. Lüsebrink, V. Schildgen, O. Schildgen
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引用次数: 0
A novel RNA virus, Thesium chinense closterovirus 1, identified by high-throughput RNA-sequencing of the parasitic plant Thesium chinense. 利用高通量RNA测序技术鉴定出一种新的中国Thesium clostervirus 1。
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_302
Chaerim Shin, Dongjin Choi, Ken Shirasu, Yasunori Ichihashi, Yoonsoo Hahn

The genome sequence of a closterovirus (genus Closterovirus, family Closteroviridae), tentatively named Thesium chinense closterovirus 1 (TcCV1), was identified by performing high-throughput RNA-sequencing of the haustoria and root tissues of Thesium chinense, a parasitic plant. The TcCV1 genome was predicted to encode nine proteins, eight of which have orthologs in previously identified closteroviruses. The TcCV1 RNA-dependent RNA polymerase (RdRp) and heat shock protein 70 homolog (Hsp70h) showed 27.8-68.2% and 23.8-55.1% amino acid identity, respectively, to orthologous proteins of known closteroviruses. The putative +1 ribosomal frameshifting site required for producing RdRp was identified as GUUUAGC with UAG stop codon and the skipped nucleotide U. Phylogenetic trees based on RdRp and Hsp70h show that TcCV1 is a novel member of the genus Closterovirus, forming a subclade with a group of known closteroviruses, including mint virus 1 and carnation necrotic fleck virus. The genome sequence of TcCV1 may be useful for studying the genome evolution of closteroviruses. Keywords: Thesium chinense closterovirus 1; Closterovirus; Closteroviridae; Thesium chinense.

通过对寄生植物Thesium chinense吸器和根组织进行高通量rna测序,鉴定了一种名为Thesium chinense clostervirus 1(暂称TcCV1)的clostervirus (clostervirus属,clostervirus科)的基因组序列。预计TcCV1基因组编码9种蛋白质,其中8种与先前鉴定的closterovirus有同源物。TcCV1 RNA依赖RNA聚合酶(RdRp)和热休克蛋白70同源物(Hsp70h)与已知closterovirus同源蛋白的氨基酸同源性分别为27.8% ~ 68.2%和23.8% ~ 55.1%。基于RdRp和Hsp70h的系统进化树表明,TcCV1是clostervirus属的一个新成员,与一组已知的clostervirus(包括薄荷病毒1和康乃馨necronecrofleck病毒)形成一个亚分支。TcCV1的基因组序列可用于研究closterovirus的基因组进化。关键词:中华葡萄球菌clostervirus 1;Closterovirus;Closteroviridae;Thesium研究方面。
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引用次数: 4
Acetylation of late expression factor 4 is crucial for the transcription and proliferation of Bombyx mori nucleopolyhedrovirus. 晚期表达因子4的乙酰化对家蚕核多角体病毒的转录和增殖至关重要。
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_109
Jonas Ngowo, Xu Gao, Yajie Zhu, Fuxiang Mao, Jihai Lei, E. Obeng, Wei Yu
Late expression factor 4 (LEF4), RNA polymerase subunit of Bombyx mori nucleopolyhedrovirus (BmNPV), plays an enzymatic role to enhance the capping of pre-mRNA of late and very late genes. Lysine acetylation is a post-translational modification process having many important functions associated with the regulation of a gene expression. Our previous study on lysine acetylome in BmNPV infected BmN cells showed that LEF 4 was acetylated at lysine 76 (K76). However, it is still unclear whether the modification of K76 residue contributes to the modulation of viral gene transcription. To elucidate the role played by acetylation or deacetylation of LEF4 K76 in the transcription of viral genes, we constructed acetylation mimicking and deacetylation mimicking mutant virus, K76Q and K76R, respectively. We then transfected BmN cells with these mutants and analyzed the level of pre-mRNA at different times. The K76R showed a significant decrease in the mRNA transcription level of vp39 and p10 genes at 48 and 72 h post-transfection, while K76Q did not show any significant changes compared with lef4-Wt. We further detected the virus titer of lef4-Wt, K76Q [et] K76R, and it was found that K76R impaired the virus infectivity ability at 72 and 96 h, while K76Q did not affect the virus infectivity. Moreover, the yeast two hybrid technique (Y2H) showed that both mutants (K76Q [et] K76R) affected the association of LEF 4 with the P47 protein. Taken together, these results indicated that acetylation modification of K76 is important for the proper transcription of late and very late genes, and the effectiveness of viral infection. Keywords: BmNPV lef4 gene; lysine acetylation, late genes transcription; BmNPV p47 gene; infectivity.
家蚕核多角体病毒(BmNPV)的RNA聚合酶亚基——晚期表达因子4 (Late expression factor 4, LEF4)在增强晚期和极晚期基因的前mrna的capping中起酶促作用。赖氨酸乙酰化是一个翻译后修饰过程,具有许多与基因表达调控相关的重要功能。我们之前对BmNPV感染的BmN细胞赖氨酸乙酰化的研究表明,lef4在赖氨酸76 (K76)处乙酰化。然而,目前尚不清楚K76残基的修饰是否参与了病毒基因转录的调节。为了阐明LEF4 K76乙酰化或去乙酰化在病毒基因转录中的作用,我们分别构建了乙酰化模拟和去乙酰化模拟突变病毒K76Q和K76R。然后我们用这些突变体转染BmN细胞,并分析不同时间pre-mRNA的水平。K76R在转染后48和72 h vp39和p10基因的mRNA转录水平显著降低,而K76Q与lef4-Wt相比没有明显变化。我们进一步检测了lef4-Wt、K76Q [et] K76R的病毒滴度,发现K76R在72和96 h时削弱了病毒的感染性,而K76Q不影响病毒的感染性。此外,酵母双杂交技术(Y2H)表明,两个突变体(K76Q [et] K76R)都影响了lef4与P47蛋白的关联。综上所述,这些结果表明,K76的乙酰化修饰对于晚期和极晚期基因的正确转录以及病毒感染的有效性至关重要。关键词:BmNPV lef4基因;赖氨酸乙酰化,晚期基因转录;BmNPV p47基因;传染性。
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引用次数: 0
Suitability of different plant species for experimental agroinfection with Plum pox virus-based expression vector for potential production of edible vaccines. 以梅痘病毒为基础的表达载体对不同植物侵染试验的适宜性及可食用疫苗的潜在生产。
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_111
Adam Achs, M. Glasa, P. Alaxin, Z. Šubr
Nine herbaceous plant species were tested for susceptibility to Plum pox virus (PPV) by Agrobacterium-mediated delivery of its infectious cDNA clone. Two of them became infected, namely spinach (local infection) and oilseed poppy (systemic infection). As a control, PPV infection was successfully established in plum seedlings following agroinfiltration, thus providing the first report of agroinfection in Prunus species. According to our results, oilseed poppy can be considered as a candidate host for the production of edible vaccines by a PPV-derived expression vector. Keywords: agroinfiltration; virus host; poppy; spinach.
采用农杆菌介导的方法,检测了9种草本植物对李痘病毒(PPV)的易感性。其中两种感染,即菠菜(局部感染)和油籽罂粟(全身感染)。作为对照,PPV侵染在灌浆后成功地在李树幼苗中建立,首次报道了李树属植物的土壤侵染。根据我们的研究结果,油籽罂粟可以被认为是ppv衍生表达载体生产可食用疫苗的候选宿主。关键词:agroinfiltration;病毒主机;罂粟花;菠菜。
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引用次数: 1
Isolation and characterization of novel Spodoptera exigua nucleopolyhedrovirus strains in Turkey and their potential for use in biological control 土耳其新夜蛾核多角体病毒株的分离、鉴定及其生物防治潜力
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_407
D. Gencer, Z. Bayramoğlu, Çetin Mutlu, I. Demir
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引用次数: 0
Genomic phylogenetic analyses of four major hand, foot and mouth disease-related enteroviruses. 四种主要手足口病相关肠道病毒的基因组系统发育分析。
IF 1.7 4区 医学 Q4 VIROLOGY Pub Date : 2022-01-01 DOI: 10.4149/av_2022_104
Yajuan Dong, Zhenzhou Wan, Shenwei Li, Jian-Hua Wang, Xia Jin, Guoying Yu, Chiyu Zhang
Enteroviruses had diverged into many types, some of which cause hand, foot and mouth disease (HFMD) in children. The predominant enterovirus types associated with HFMD are EVA71, CVA16, CVA6 and CVA10. Four enterovirus types were classified into subtypes based on VP1 sequences. However, the phylogenetics of these enteroviruses is rarely concerned at the genomic level. In this study, we performed the phylogenetic analyses of the EVA71, CVA16, CVA6 and CVA10 using available full-length genomic sequences. We found that the topologies of phylogenetic trees of full-length genomic sequences and VP1 sequences were almost consistent, except few subtypes of EVA71 and CVA10. The mean genetic divergence was 15.8-27% between subtypes and less than 12% within subtypes/sub-subtypes at genomic level. Comparison of phylogenetic topologies between genomic and VP1 sequences helped us to identify two new EVA71 inter-subtype recombinants RF01_CC4 and RF02_CC4. Furthermore, EVA71 subtypes C1 and C2 and CVA10 subtype D were found to originate through inter-subtype recombination. The genomic reference sequences of these enteroviruses are provided here for subtyping. The results provide important insights into the understanding of the evolution and epidemiology of the four enteroviruses. Keywords: enterovirus; hand; foot and mouth disease; classification; genetic distance; recombination.
肠病毒已经分化成许多类型,其中一些会导致儿童手足口病。与手足口病相关的主要肠道病毒类型为EVA71、CVA16、CVA6和CVA10。根据VP1序列将4种肠道病毒分为不同的亚型。然而,这些肠道病毒的系统发育很少在基因组水平上得到关注。在这项研究中,我们利用现有的全基因组序列对EVA71、CVA16、CVA6和CVA10进行了系统发育分析。研究发现,除了EVA71和CVA10的少数亚型外,VP1全长基因组序列与VP1序列的系统发育树拓扑结构基本一致。亚型间遗传差异均值为15.8 ~ 27%,亚型/亚亚型间遗传差异均值小于12%。通过比较基因组序列和VP1序列的系统发育拓扑结构,我们鉴定出两个新的EVA71亚型间重组基因RF01_CC4和RF02_CC4。此外,发现EVA71亚型C1和C2以及CVA10亚型D是通过亚型间重组产生的。这里提供了这些肠道病毒的基因组参考序列用于分型。这些结果为了解这四种肠道病毒的进化和流行病学提供了重要的见解。关键词:肠病毒;手;口蹄疫;分类;遗传距离;重组。
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引用次数: 0
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Acta virologica
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