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      Characterisation of divergent flavivirus NS3 and NS5 protein sequences detected in Rhipicephalus microplus ticks from Brazil

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          Abstract

          Transcripts similar to those that encode the nonstructural (NS) proteins NS3 and NS5 from flaviviruses were found in a salivary gland (SG) complementary DNA (cDNA) library from the cattle tick Rhipicephalus microplus. Tick extracts were cultured with cells to enable the isolation of viruses capable of replicating in cultured invertebrate and vertebrate cells. Deep sequencing of the viral RNA isolated from culture supernatants provided the complete coding sequences for the NS3 and NS5 proteins and their molecular characterisation confirmed similarity with the NS3 and NS5 sequences from other flaviviruses. Despite this similarity, phylogenetic analyses revealed that this potentially novel virus may be a highly divergent member of the genus Flavivirus. Interestingly, we detected the divergent NS3 and NS5 sequences in ticks collected from several dairy farms widely distributed throughout three regions of Brazil. This is the first report of flavivirus-like transcripts in R. microplus ticks. This novel virus is a potential arbovirus because it replicated in arthropod and mammalian cells; furthermore, it was detected in a cDNA library from tick SGs and therefore may be present in tick saliva. It is important to determine whether and by what means this potential virus is transmissible and to monitor the virus as a potential emerging tick-borne zoonotic pathogen.

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          Most cited references62

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          SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler

          Background There is a rapidly increasing amount of de novo genome assembly using next-generation sequencing (NGS) short reads; however, several big challenges remain to be overcome in order for this to be efficient and accurate. SOAPdenovo has been successfully applied to assemble many published genomes, but it still needs improvement in continuity, accuracy and coverage, especially in repeat regions. Findings To overcome these challenges, we have developed its successor, SOAPdenovo2, which has the advantage of a new algorithm design that reduces memory consumption in graph construction, resolves more repeat regions in contig assembly, increases coverage and length in scaffold construction, improves gap closing, and optimizes for large genome. Conclusions Benchmark using the Assemblathon1 and GAGE datasets showed that SOAPdenovo2 greatly surpasses its predecessor SOAPdenovo and is competitive to other assemblers on both assembly length and accuracy. We also provide an updated assembly version of the 2008 Asian (YH) genome using SOAPdenovo2. Here, the contig and scaffold N50 of the YH genome were ~20.9 kbp and ~22 Mbp, respectively, which is 3-fold and 50-fold longer than the first published version. The genome coverage increased from 81.16% to 93.91%, and memory consumption was ~2/3 lower during the point of largest memory consumption.
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            Precision Farming: Technologies and Information as Risk-Reduction Tools

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              The codon Adaptation Index--a measure of directional synonymous codon usage bias, and its potential applications.

              P. Sharp, W Li (1987)
              A simple, effective measure of synonymous codon usage bias, the Codon Adaptation Index, is detailed. The index uses a reference set of highly expressed genes from a species to assess the relative merits of each codon, and a score for a gene is calculated from the frequency of use of all codons in that gene. The index assesses the extent to which selection has been effective in moulding the pattern of codon usage. In that respect it is useful for predicting the level of expression of a gene, for assessing the adaptation of viral genes to their hosts, and for making comparisons of codon usage in different organisms. The index may also give an approximate indication of the likely success of heterologous gene expression.
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                Author and article information

                Journal
                Mem Inst Oswaldo Cruz
                Mem. Inst. Oswaldo Cruz
                Memórias do Instituto Oswaldo Cruz
                Instituto Oswaldo Cruz, Ministério da Saúde
                0074-0276
                1678-8060
                10 October 2013
                February 2014
                : 109
                : 1
                : 38-50
                Affiliations
                [1 ] Faculdade de Medicina de Ribeirão Preto
                [2 ] National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD, USA
                [3 ] Faculdade de Tecnologia, Araçatuba, SP, Brasil
                [4 ] Vallée SA, São Paulo, SP, Brasil
                [5 ] Laticínios Vale dos Buritis, Buritis, MG, Brasil
                [6 ] Vetmais Medicina Animal, Campinas, SP, Brasil
                [7 ] Universidade de São Paulo, Escola de Enfermagem de Ribeirão Preto, Ribeirão Preto, SP, Brasil, Escola de Enfermagem de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil
                Author notes
                [+ ] Corresponding author: imsantos@fmrp.usp.br
                Article
                10.1590/0074-0276130166
                4005522
                24626302
                327434bd-9d32-4a4e-b83d-4a6e46da551c

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 25 March 2013
                : 16 August 2013
                Page count
                Figures: 7, Tables: 2, References: 69, Pages: 13
                Funding
                Funded by: FAPESP
                Award ID: 2004/09992-7
                Funded by: FAPESP
                Award ID: 2009/53645-3
                Funded by: FAPESP
                Award ID: 2012/06374-7
                Funded by: FAPESP
                Award ID: 2006/54041-6
                Funded by: FAPESP
                Award ID: 2007/59357-4
                Funded by: FAPESP
                Award ID: 2009/51212-2
                Funded by: CNPq
                Award ID: 559603/2009-6
                Funded by: CNPq
                Award ID: 471946/2010-9
                Funded by: CNPq
                Award ID: 420067/2005-1
                Funded by: CNPq
                Award ID: 505810/2004-2
                Funded by: CNPq
                Award ID: 143354/2008-6
                Categories
                Articles

                rhipicephalus microplus,tick,ns3,ns5,flavivirus
                rhipicephalus microplus, tick, ns3, ns5, flavivirus

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