930 resultados para Clones recombinantes


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The red gum lerp psyllid Glycaspis brimblecombei Moore (Hemiptera: Psyllidae) was first recorded in Brazil in 2003 and since then has caused serious damage in Eucalyptus forests. The damage is mainly due to defoliation, sooty mold and dieback, thus requiring the development of efficient management strategies. The settling and ovipositional preference of G. brimblecombei by species and clones of Eucalyptus were evaluated in free- and no-choice tests. Preliminary tests were done to determine the density of psyllid couples that produced the greatest densities of eggs, leaf position selection for oviposition, and whether oviposition was affected by preimaginal conditioning. Apical leaves were used more frequently for oviposition by the psyllid, and the number of eggs decreased from the apex to the base. The densities of 10 and 15 psyllid couples per seedling produced the highest oviposition. There was no evidence of preimaginal conditioning. In the free-choice test, E. grandis, E. urophylla, VM-1, I-144, C-219 and H-13 were less attractive to G. brimblecombei adults, with H-13, E. grandis and E. urophylla having the least oviposition. In the no-choice test, E. urophylla, GG-100 and E. grandis also demonstrated a lower oviposition, but E. camaldulensis and 3025 were highly susceptible. The low preference for the genotypes E. grandis and E. urophylla suggests the occurrence of a non-preference type resistance against the red gum lerp psyllid. Our results can be utilized for the development of management programs for G. brimblecombei in Eucalyptus forests. © 2012 Springer Science + Business Media B.V.

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Background:Hepatitis C is a disease spread throughout the world. Hepatitis C virus (HCV), the etiological agent of this disease, is a single-stranded positive RNA virus. Its genome encodes a single precursor protein that yields ten proteins after processing. NS5A, one of the non-structural viral proteins, is most associated with interferon-based therapy response, the approved treatment for hepatitis C in Brazil. HCV has a high mutation rate and therefore high variability, which may be important for evading the immune system and response to therapy. The aim of this study was to analyze the evolution of NS5A quasispecies before, during, and after treatment in patients infected with HCV genotype 3a who presented different therapy responses.Methods:Viral RNA was extracted, cDNA was synthesized, the NS5A region was amplified and cloned, and 15 clones from each time-point were sequenced. The sequences were analyzed for evolutionary history, genetic diversity and selection.Results:This analysis shows that the viral population that persists after treatment for most non-responder patients is present in before-treatment samples, suggesting it is adapted to evade treatment. In contrast, the population found in before treatment samples from most end-of-treatment responder patients either are selected out or appears in low frequency after relapse, therefore changing the population structure. The exceptions illustrate the uniqueness of the evolutionary process, and therefore the treatment resistance process, in each patient.Conclusion:Although evolutionary behavior throughout treatment showed that each patient presented different population dynamics unrelated to therapy outcome, it seems that the viral population from non-responders that resists the treatment already had strains that could evade therapy before it started. © 2013 Bittar et al.

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Even though community-acquired methicillin resistant Staphylococcus aureus (CA-MRSA) was described a decade ago, reports from Brazil are scarce and cases occurred in large urban centers. We report MRSA sepsis in a 16-year-old male from a small town and who had no history of exposure to healthcare or recent travel. After trauma during a soccer match, he presented swelling in the right thigh, which evolved in a month to cellulitis complicated by local abscess, orchitis and pneumonia. The patient presented severe sepsis, with fever and respiratory failure. Laboratory findings included blood leukocyte counts above 40,000/mm3 and thrombocytopenia. He was submitted to mechanical ventilation and therapy with vancomycin and imipenem. He had a slow but favorable response to therapy and was discharged after six weeks of hospitalization. MRSA grew from blood cultures and respiratory aspirates obtained before antimicrobial therapy. The isolate belonged to sequence type 5, spa type t311, harbored SCCmec type IV and genes for Panton-Valentine leukocidin and Enterotoxin A. The pulsed-field gel electrophoresis pattern was distinct from North American classic CA-MRSA clones. However, the sequence type and the spa type revealed that the clone belong to the same clonal complex isolated in Argentina. This is the first CA-MRSA infection reported in that region, with significant epidemiologic and clinical implications. © 2013 Elsevier Inc.

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Background: Transposable elements (TEs) have the potential to produce broad changes in the genomes of their hosts, acting as a type of evolutionary toolbox and generating a collection of new regulatory and coding sequences. Several TE classes have been studied in Neotropical cichlids; however, the information gained from these studies is restricted to the physical chromosome mapping, whereas the genetic diversity of the TEs remains unknown. Therefore, the genomic organization of the non-LTR retrotransposons Rex1, Rex3, and Rex6 in five Amazonian cichlid species was evaluated using physical chromosome mapping and DNA sequencing to provide information about the role of TEs in the evolution of cichlid genomes. Results: Physical mapping revealed abundant TE clusters dispersed throughout the chromosomes. Furthermore, several species showed conspicuous clusters accumulation in the centromeric and terminal portions of the chromosomes. These TE chromosomal sites are associated with both heterochromatic and euchromatic regions. A higher number of Rex1 clusters were observed among the derived species. The Rex1 and Rex3 nucleotide sequences were more conserved in the basal species than in the derived species; however, this pattern was not observed in Rex6. In addition, it was possible to observe conserved blocks corresponding to the reverse transcriptase fragment of the Rex1 and Rex3 clones and to the endonuclease of Rex6. Conclusion: Our data showed no congruence between the Bayesian trees generated for Rex1, Rex3 and Rex6 of cichlid species and phylogenetic hypothesis described for the group. Rex1 and Rex3 nucleotide sequences were more conserved in the basal species whereas Rex6 exhibited high substitution rates in both basal and derived species. The distribution of Rex elements in cichlid genomes suggests that such elements are under the action of evolutionary mechanisms that lead to their accumulation in particular chromosome regions, mostly in heterochromatins. © 2013 Schneider et al.; licensee BioMed Central Ltd.

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The major histocompatibility complex (MHC) in mammals codes for antigen-presenting proteins. For this reason, the MHC is of great importance for immune function and animal health. Previous studies revealed this gene-dense and polymorphic region in river buffalo to be on the short arm of chromosome 2, which is homologous to cattle chromosome 23. Using cattle-derived STS markers and a river buffalo radiation hybrid (RH) panel (BBURH5000), we generated a high-resolution RH map of the river buffalo MHC region. The buffalo MHC RH map (cR5000) was aligned with the cattle MHC RH map (cR 12000) to compare gene order. The buffalo MHC had similar organization to the cattle MHC, with class II genes distributed in two segments, class IIa and class IIb. Class IIa was closely associated with the class I and class III regions, and class IIb was a separate cluster. A total of 53 markers were distributed into two linkage groups based on a two-point LOD score threshold of ≥8. The first linkage group included 32 markers from class IIa, class I and class III. The second linkage group included 21 markers from class IIb. Bacterial artificial chromosome clones for seven loci were mapped by fluorescence in situ hybridization on metaphase chromosomes using single- and double-color hybridizations. The order of cytogenetically mapped markers in the region corroborated the physical order of markers obtained from the RH map and served as anchor points to align and orient the linkage groups. © 2012 The Authors, Animal Genetics © 2012 Stichting International Foundation for Animal Genetics.

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Methicillin-resistant Staphylococcus aureus (MRSA) poses a threat for patients in burn units. Studies that mix epidemiological designs with molecular typing may contribute to the development of strategies for MRSA control. We conducted a study including: molecular characterization of Staphylococcal Chromosome Cassette mecA (SCCmec), strain typing with pulsed field gel electrophoresis (PFGE) and detection of virulence genes, altogether with a case-case-control study that assessed risk factors for MRSA and for methicillin-susceptible S. aureus (MSSA), using S. aureus negative patients as controls. Strains were collected from clinical and surveillance cultures from October 2006 through March 2009. MRSA was isolated from 96 patients. Most isolates (94.8%) harbored SCCmec type III. SCCmec type IV was identified in isolates from four patients. In only one case it could be epidemiologically characterized as community-associated. PFGE typing identified 36 coexisting MRSA clones. When compared to MSSA (38 isolates), MRSA isolates were more likely to harbor two virulence genes: tst and lukPV. Previous stay in other hospital and admission to Intensive Care Unit were independent risk factors for both MRSA and MSSA, while the number of burn wound excisions was significantly related with the former (OR = 6.80, 95%CI = 3.54-13.07). In conclusion, our study found polyclonal endemicity of MRSA in a burn unit, possibly related to importing of strains from other hospitals. Also, it pointed out to a role of surgical procedures in the dissemination of MRSA strains. © 2013 Elsevier Ltd and ISBI. All rights reserved.

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Fungi constitute an important part of the soil ecosystem, playing key roles in decomposition, cycling processes, and biotic interactions. Molecular methods have been used to assess fungal communities giving a more realistic view of their diversity. For this purpose, total DNA was extracted from bulk soils cultivated with tomato (STC), vegetables (SHC), and native forest (SMS) from three sites of the Taquara Branca river basin in Sumaré County, São Paulo State, Brazil. This metagenomic DNA was used as a template to amplify fungal 18S rDNA sequences, and libraries were constructed in Escherichia coli by cloning PCR products. The plasmid inserts were sequenced and compared to known rDNA sequences in the GenBank database. Of the sequenced clones, 22 were obtained from the SMS sample, 18 from the SHC sample, and 6 from the STC sample. Although most of the clone sequences did not match the sequences present in the database, individual amplified sequences matched with Glomeromycota (SMS), Fungi incertae sedis (SMS), and Neocallimastigomycota (SHC). Most of the sequences from the amplified taxa represent uncultured fungi. The molecular analysis of variance (AMOVA) indicated that fluctuations observed of haplotypes in the composition may be related to herbicide application. © 2013 Silvana Pompéia Val-Moraes et al.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

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Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Pós-graduação em Ciência Florestal - FCA