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Zhao H Yu XD Zhang XY Jiang T Hong WX Yu M Hu FY Zhu SY Qin ED Deng YQ Qin CF Zhang FC 《Journal of virology》2012,86(12):7021-7022
Here we report the first complete genome sequence of a dengue virus serotype 4 genotype II strain, GZ30, isolated in Guangzhou, Guangdong Province, China, in 2010. The sequence information provided herein will help us to understand the molecular epidemiology of dengue virus and predict the risk of severe diseases in mainland China. 相似文献
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Naveca FG Souza VC Silva GA Maito RM Granja F Siqueira T Acosta PO 《Journal of virology》2012,86(3):1897-1898
Dengue is the most important arboviral disease worldwide. We report the complete genome sequence of a dengue virus serotype 4, genotype II strain isolated in 2010 from a patient with classical dengue fever in Boa Vista, Roraima, Brazil. 相似文献
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Jefferson José da Silva Santos Tereza Magalh?es José Valter Joaquim Silva Junior Andréa Nazaré Monteiro Rangel da Silva Marli Tenório Cordeiro Laura Helena Vega Gonzales Gil 《Memórias do Instituto Oswaldo Cruz》2015,110(5):677-683
Full-length dengue virus (DENV) cDNA clones are an invaluable tool for many studies,
including those on the development of attenuated or chimeric vaccines and on
host-virus interactions. Furthermore, the importance of low passage DENV infectious
clones should be highlighted, as these may harbour critical and unique
strain-specific viral components from field-circulating isolates. The successful
construction of a functional Brazilian low passage DENV serotype 2 full-length clone
through homologous recombination reported here supports the use of a strategy that
has been shown to be highly useful by our group for the development of flavivirus
infectious clones and replicons. 相似文献
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ZHAO Hui ZHAO Ling Zhai JIANG Tao LI Xiao Feng FAN Hang HONG Wen Xin ZHANG Yu ZHU Qin YE Qing TONG Yi Gang CAO Wu Chun ZHANG Fu Chun QIN Cheng Feng 《中国科学:生命科学英文版》2014,(12)
Dengue has been well recognized as a global public health threat,but only sporadic epidemics and imported cases were reported in recent decades in China.Since July 2014,an unexpected large dengue outbreak has occurred in Guangdong province,China,resulting in more than 40000 patients including six deaths.To clarify and characterize the causative agent of this outbreak,the acute phase serum from a patient diagnosed with severe dengue was subjected to virus isolation and high-throughput sequencing(HTS).Traditional real-time RT-PCR and HTS with Ion Torrent PGM detected the presence of dengue virus serotype 2(DENV-2).A clinical DENV-2 isolate GZ05/2014 was obtained by culturing the patient serum in mosquito C6/36 cells.The complete genome of GZ05/2014 was determined and deposited in Gen Bank under the access number KP012546.Phylogenetic analysis based on the complete envelope gene showed that the newly DENV-2 isolate belonged to Cosmopolitan genotype and clustered closely with other Guangdong strains isolated in the past decade.No amino acid mutations that are obviously known to increase virulence or replication were identified throughout the genome of GZ05/2014.The high homology of Guangdong DENV-2 strains indicated the possibility of establishment of local DENV-2 circulation in Guangdong,China.These results help clarify the origin of this epidemic and predict the future status of dengue in China. 相似文献
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The plasticity of viral plus strand RNA genomes is fundamental for the multiple functions of these molecules. Local and long-range RNA-RNA interactions provide the scaffold for interacting proteins of the translation, replication, and encapsidation machinery. Using dengue virus as a model, we investigated the relevance of the interplay between two alternative conformations of the viral genome during replication. Flaviviruses require long-range RNA-RNA interactions and genome cyclization for RNA synthesis. Here, we define a sequence present in the viral 3'UTR that overlaps two mutually exclusive structures. This sequence can form an extended duplex by long-range 5'-3' interactions in the circular conformation of the RNA or fold locally into a small hairpin (sHP) in the linear form of the genome. A mutational analysis of the sHP structure revealed an absolute requirement of this element for viral viability, suggesting the need of a linear conformation of the genome. Viral RNA replication showed high vulnerability to changes that alter the balance between circular and linear forms of the RNA. Mutations that shift the equilibrium toward the circular or the linear conformation of the genome spontaneously revert to sequences with different mutations that tend to restore the relative stability of the two competing structures. We propose a model in which the viral genome exists in at least two alternative conformations and the balance between these two states is critical for infectivity. 相似文献
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Biological markers are normally used to evaluate the candidate of live-attenuated dengue vaccines. D3V 16562 Vero 23 and D3V
16562 Vero 33 which were derivatives of D3V 16562, parental strain, showed the similar biological data. We used molecular
techniques and computational tools to evaluate these derivatives. The nucleotide and amino acid sequences of the derivatives were
compared to their parent. The secondary structures of untranslated regions and B-cell epitopes were predicted. The results showed
that nucleotide substitutions mostly occurred in NS5 and NS5 of V2 was unusual because of amino acid change at 3349 (tryptophan
→stop codon). The nucleotide substitutions in 5''UTR, prM, E, NS1, NS2A, NS3, and 3''UTR were 4, 1, 2, 2, 1, 3, and 2, respectively.
The secondary structure of 5''UTR of V2 was different from P and V1. The secondary structure of 3''UTR of V2 was similar to P and
certainly distinct from V1. Furthermore, B-cell epitopes prediction revealed that there were 21 epitopes of envelope and the
interesting epitope was at position 297-309 because it was in domain III in which the neutralizing antibody is induced. For this
study, the attenuation of derivatives was caused by the nucleotide substitutions in 5''UTR, 3''UTR, and NS5 regions. The genotypic
data and B-cell epitope make the derivatives attractive for the chimeric and peptide DENV vaccine development. 相似文献
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目的构建以乙型脑炎病毒(Japanese encephalitis virus,JEV)疫苗株SA14-14-2为基因骨架的乙脑/登革4型嵌合病毒,并分析该嵌合病毒对小鼠的神经毒力。方法通过重叠PCR方法扩增含有登革病毒4型(DENV-4)H241株pr ME基因序列和乙型脑炎病毒疫苗株SA14-14-2的NS1蛋白前177个核苷酸的融合片段,用Nar I和Bgl II双酶切后替换乙型脑炎病毒疫苗株SA14-14-2全长克隆中的相应区域,构建成乙脑/登革4型嵌合全长克隆,通过体外转录和转染BHK21细胞获得嵌合病毒(JEV/DENV-4 chimeric virus,JD4)。通过测定嵌合病毒JD4和2个母本株JEV SA14-14-2株及DENV-4 H241株蚀斑大小、小鼠脑内神经毒力和皮下感染入脑能力、乳鼠脑内神经毒力,比较JD4和母本株之间的差异。通过将JD4在原代地鼠肾(primary hamster kidney,PHK)细胞传代30次,分析传代后嵌合病毒的神经毒力是否减弱及减弱的程度。结果测序结果表明,构建的嵌合病毒JD4基因组序列和预期一致,没有产生新的位点突变。JD4蚀斑较SA14-14-2明显偏小,但和DENV-4 H241株没有明显区别。JD4对3周龄小鼠具有较强的脑内神经毒力,和母本株DENV-4 H241没有差异,对小鼠没有神经侵袭力。乳鼠实验结果表明,嵌合病毒JD4脑内神经毒力虽然略低于母本株DENV-4 H241,但两者之间没有明显差异,都明显强于乙脑疫苗株SA14-14-2。在PHK细胞传代30次后,小鼠神经毒力虽然有所减低,但并不明显。结论成功构建了嵌合病毒JD4,通过测定并比较JD4与母本株的蚀斑特征、小鼠及乳鼠神经毒力等试验,为分析登革疫苗候选株安全性研究奠定了基础。 相似文献
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Alex Pauvolid-Corrêa Raquel Soares Juliano Zilca Campos Jason Velez Rita Maria Ribeiro Nogueira Nicholas Komar 《Memórias do Instituto Oswaldo Cruz》2015,110(1):125-133
The Pantanal hosts diverse wildlife species and therefore is a hotspot for arbovirus
studies in South America. A serosurvey for Mayaro virus (MAYV), eastern (EEEV),
western (WEEV) and Venezuelan (VEEV) equine encephalitis viruses was conducted with
237 sheep, 87 free-ranging caimans and 748 equids, including 37 collected from a
ranch where a neurologic disorder outbreak had been recently reported. Sera were
tested for specific viral antibodies using plaque-reduction neutralisation test. From
a total of 748 equids, of which 264 were immunised with vaccine composed of EEEV and
WEEV and 484 had no history of immunisation, 10 (1.3%) were seropositive for MAYV and
two (0.3%) for VEEV using criteria of a ≥ 4-fold antibody titre difference. Among the
484 equids without history of immunisation, 48 (9.9%) were seropositive for EEEV and
four (0.8%) for WEEV using the same criteria. Among the sheep, five were sero-
positive for equine encephalitis alphaviruses, with one (0.4%) for EEEV, one (0.4%)
for WEEV and three (1.3%) for VEEV. Regarding free-ranging caimans, one (1.1%) and
three (3.4%), respectively, had low titres for neutralising antibodies to VEEV and
undetermined alphaviruses. The neurological disorder outbreak could not be linked to
the alphaviruses tested. Our findings represent strong evidence that MAYV and all
equine encephalitis alphaviruses circulated in the Pantanal. 相似文献
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Complete genome sequence of an aerobic thermoacidophilic crenarchaeon, Sulfolobus tokodaii strain7. 总被引:7,自引:0,他引:7
Y Kawarabayasi Y Hino H Horikawa K Jin-no M Takahashi M Sekine S Baba A Ankai H Kosugi A Hosoyama S Fukui Y Nagai K Nishijima R Otsuka H Nakazawa M Takamiya Y Kato T Yoshizawa T Tanaka Y Kudoh J Yamazaki N Kushida A Oguchi K Aoki S Masuda M Yanagii M Nishimura A Yamagishi T Oshima H Kikuchi 《DNA research》2001,8(4):123-140
The complete genomic sequence of an aerobic thermoacidophilic crenarchaeon, Sulfolobus tokodaii strain7 which optimally grows at 80 degrees C, at low pH, and under aerobic conditions, has been determined by the whole genome shotgun method with slight modifications. The genomic size was 2,694,756 bp long and the G + C content was 32.8%. The following RNA-coding genes were identified: a single 16S-23S rRNA cluster, one 5S rRNA gene and 46 tRNA genes (including 24 intron-containing tRNA genes). The repetitive sequences identified were SR-type repetitive sequences, long dispersed-type repetitive sequences and Tn-like repetitive elements. The genome contained 2826 potential protein-coding regions (open reading frames, ORFs). By similarity search against public databases, 911 (32.2%) ORFs were related to functional assigned genes, 921 (32.6%) were related to conserved ORFs of unknown function, 145 (5.1%) contained some motifs, and remaining 849 (30.0%) did not show any significant similarity to the registered sequences. The ORFs with functional assignments included the candidate genes involved in sulfide metabolism, the TCA cycle and the respiratory chain. Sequence comparison provided evidence suggesting the integration of plasmid, rearrangement of genomic structure, and duplication of genomic regions that may be responsible for the larger genomic size of the S. tokodaii strain7 genome. The genome contained eukaryote-type genes which were not identified in other archaea and lacked the CCA sequence in the tRNA genes. The result suggests that this strain is closer to eukaryotes among the archaea strains so far sequenced. The data presented in this paper are also available on the internet homepage (http://www.bio.nite.go.jp/E-home/genome_list-e.html/). 相似文献
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我国登革 4型病毒 B5株基因组全序列的测定及分析(英文) 总被引:2,自引:0,他引:2
对我国登革 4型病毒 B5株 (D4- B5)基因组进行全序列测定及分析 ,为研究病毒基因组结构与功能的关系及研制新型登革疫苗奠定基础 .根据登革 4型病毒 81 4669株的序列设计特异引物 ,通过 RT- PCR扩增出 D4- B5株不同长度的片段 ,分别克隆到 p GEM- T载体 ,将挑取的阳性克隆进行 PCR、酶切鉴定及序列测定 .结果显示 ,D4- B5株的基因组全长 1 0 665nt,5′和 3′非编码区分别为 1 0 1 nt和 40 3nt,中间一个长 1 0 1 61 nt的开放读码框架 ,编码 3387个氨基酸 .与 D4- 81 4669株比较 ,两者核苷酸序列同源性为 93.0 8% ,氨基酸序列同源性为 96.58% .D4- B5株的基因组全序列与 D4- 81 4669株类似 ,但也有较大差异 .同源进化分析表明 ,D4- B5株的基因型为 型 ,与登革 4型病毒菲律宾分离株亲缘关系较近 .这是首次报道的我国登革 4型病毒分离株基因组全序列 ,对研究病毒基因组结构与功能的关系 ,探讨我国毒株的地理来源及研制适合我国人群的新型登革疫苗具有一定的意义 . 相似文献
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Induction of multiple cytotoxic T lymphocyte responses in mice by a multiepitope DNA vaccine against dengue virus serotype 1 下载免费PDF全文
Xin Yu Chen De Zhou Li Xiao Zhi Zhong Bokun Chen Zhi Liang Duan Jin Sheng Wen 《Microbiology and immunology》2016,60(12):835-845
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一株A型口蹄疫流行毒株全序列的测定及其全长感染性克隆的构建 总被引:1,自引:0,他引:1
【目的】测定一株A型口蹄疫流行毒株的全基因组序列,并构建其全长感染性克隆。【方法】参照已公布的A型口蹄疫病毒序列设计引物,将分离的口蹄疫病毒株A/Sea-97/CHA/2014全基因组分为4个重叠的片段进行RT-PCR扩增,并对其进行序列测定与分析。利用酶切连接法将4个基因片段依次克隆至p Blue Script SKhdv载体中,构建该流行毒株的全长c DNA克隆p QAHN。pQAHN经NotⅠ线性化后转染表达T7 RNA聚合酶的BSR/T7细胞,拯救病毒。【结果】口蹄疫病毒全基因组序列测定结果表明该毒株基因组全长8 171 bp[不包括poly(C)区段和poly(A)尾巴],开放阅读框为6 996 bp,编码2 332个氨基酸,5′和3′非编码区分别为1 091 bp和95 bp。VP1系统发生树分析表明该毒株与A/GDMM/CHA/2013毒株亲缘关系最近,相似性为99.1%。线化全长质粒转染BSR/T7细胞68 h后可观察到典型的细胞病变。拯救病毒的间接免疫荧光、RT-PCR和序列测定结果表明成功拯救出了具有感染性的FMDV。拯救病毒与亲本病毒的噬斑表型及生长曲线试验表明二者具有相似的生长表型和增殖能力。【结论】该研究为我国口蹄疫病原生态分布、分子流行病学调查以及A型FMD新型疫苗的研究提供了有益的材料。 相似文献
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Bruno Gabriel N Andrade Michel F Abanto Marin Diego Duque Cambuy Erica Louren?o Fonseca Nadjla Ferreira Souza Ana Carolina P Vicente 《Memórias do Instituto Oswaldo Cruz》2014,109(7):972-974
There has been a resurgence in the number of pertussis cases in Brazil and around theworld. Here, the genome of a clinical Bordetella pertussis strain (Bz181) that wasrecently isolated in Brazil is reported. Analysis of the virulence-associated genesdefining the pre- and post-vaccination lineages revealed the presence of theprn2-ptxS1A-fim3B-ptxP3 allelic profile in Bz181, which is characteristic of thecurrent pandemic lineage. A putative metallo-β-lactamase gene presenting all of theconserved zinc-binding motifs that characterise the catalytic site was identified, inaddition to a multidrug efflux pump of the RND family that could confer resistance toerythromycin, which is the antibiotic of choice for treating pertussis disease. 相似文献
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Three new species of orchid bees are described and figured from the Amazon and Atlantic forests of Brazil. Euglossa clausi
sp. n., Euglossa moratoi
sp. n., and Euglossa pepei
sp. n. are distinguished from their close congeners Euglossa crassipunctata Moure, Euglossa parvula Dressler, and Euglossa sapphirina Moure, previously placed in the subgenus Euglossa (Glossurella) Dressler, 1982, a demonstrably paraphyletic assemblage requiring serious reconsideration. Their affinities with related species are discussed and pertinent characters are figured. 相似文献
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The nucleotide sequence of the entire chloroplast genome (150,519 bp) of a legume, Lotus japonicus, has been determined. The circular double-stranded DNA contains a pair of inverted repeats of 25,156 bp which are separated by a small and a large single copy region of 18,271 bp and 81,936 bp, respectively. A total of 84 predicted protein-coding genes including 7 genes duplicated in the inverted repeat regions, 4 ribosomal RNA genes and 37 tRNA genes (30 gene species) representing 20 amino acids species were assigned on the genome based on similarity to genes previously identified in other chloroplasts. All the predicted genes were conserved among dicot plants except that rpl22, a gene encoding chloroplast ribosomal protein CL22, was missing in L. japonicus. Inversion of a 51-kb segment spanning rbcL to rpsl6 (positions 5161-56,176) in the large single copy region was observed in the chloroplast genome of L. japonicus. The sequence data and gene information are available on our World Wide Web database at http://www.kazusa.or.jp/en/plant/database.html. 相似文献
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Ravin NV Mardanova ES Kotlyarov RY Novikov VK Atabekov JG Skryabin KG 《Biochemistry. Biokhimii?a》2008,73(1):44-49
The complete nucleotide sequence of the genome of a new potato virus X (PVX) strain Tula isolated by us has been determined. Based on comparison of the PVX Tula nucleotide sequence with the sequences of 12 other PVX strains, this strain was assigned to the European cluster of PVX strains. Phylogenetic analysis revealed the same phylogeny for both full genome sequences and nucleotide sequences of polymerase and coat protein genes, suggesting that the PVX evolution did not involve recombination between different strains. The full-size cDNA copy of the PVX Tula genome was cloned and the accumulation of the viral coat protein in infected Nicotiana benthamiana was shown to be about twofold higher than for the PVX strain UK3. Based on the PVX Tula genome, a new vector which contained the target gene instead of the removed triple transport gene block and the coat protein gene has been constructed for expression of target proteins in plants. The productivity of the new vector was about 1.5-2-fold higher than the productivity of the vector of the same structure based on the standard PVX strain genome. The new viral vector can be used for superproduction of recombinant proteins in plants. 相似文献
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《Standards in genomic sciences》2013,8(2):188-197
Streptococcus agalactiae (Lancefield group B; GBS) is the causative agent of meningoencephalitis in fish, mastitis in cows, and neonatal sepsis in humans. Meningoencephalitis is a major health problem for tilapia farming and is responsible for high economic losses worldwide. Despite its importance, the genomic characteristics and the main molecular mechanisms involved in virulence of S. agalactiae isolated from fish are still poorly understood. Here, we present the genomic features of the 1,820,886 bp long complete genome sequence of S. agalactiae SA20-06 isolated from a meningoencephalitis outbreak in Nile tilapia (Oreochromis niloticus) from Brazil, and its annotation, consisting of 1,710 protein-coding genes (excluding pseudogenes), 7 rRNA operons, 79 tRNA genes and 62 pseudogenes. 相似文献