Navegando por Palavras-chave "Sequence"

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    Mapping and validation of xanthomonas citri subsp citri genes regulated by putative plant-inducible promoter box (pip-box)
    (Dove Medical Press Ltd, 2016) Carvalho, F. M. S.; Oliveira, J. C. F. [UNIFESP]; Laia, M. L.; Jacob, T. R.; Ferreira, R. M.; Ferro, M. I. T.; Tezza, R. I. D.; Zingaretti, S. M.; Silva, C. F.; Ferro, J. A.
    Citrus canker, caused by the Gram-negative bacterium Xanthomonas citri subsp citri (Xac), is a major disease affecting citriculture worldwide, because of the susceptibility of the host and the lack of efficient control methods. Previous studies have reported that some genes of phytopathogenic bacteria possess a consensus nucleotide sequence (TTCGC...N15...TTCGC) designated the "plant-inducible-promoter box" (PIP box) located in the promoter region, which is responsible for activating the expression of pathogenicity and virulence factors when the pathogen is in contact with the host plant. In this study, we mapped and investigated the expression of 104 Xac genes associated with the PIP box sequences using a macroarray analysis. Xac gene expression was observed during in vitro (Xac grown for 12 or 20 h in XAM1 induction medium) or in vivo (bacteria grown in orange leaves for 3 to 5 days) infection conditions. Xac grown in non-induction NB liquid medium was used as the control. cDNA was isolated from bacteria grown under the different conditions and hybridized to the macroarray, and 32 genes differentially expressed during the infection period (in vitro or in vivo induction) were identified. The macroarray results were validated for some of the genes through semi-quantitative RT-PCR, and the functionality of the PIP box-containing promoter was demonstrated by activating beta-glucuronidase reporter gene activity by the PIP box-containing promoter region during Xac-citrus host interaction.
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    Molecular identification of Histoplasma capsulatum using rolling circle amplification
    (Wiley-Blackwell, 2016) Furuie, Jason L.; Sun, Jiufeng; do Nascimento, Mariana M. F.; Gomes, Renata R.; Waculicz-Andrade, Caroline E.; Sessegolo, Gisele C.; Rodrigues, Anderson M. [UNIFESP]; Galvao-Dias, Maria A.; de Camargo, Zoilo P. [UNIFESP]; Queiroz-Telles, Flavio; Najafzadeh, Mohammad J.; de Hoog, Sybren G.; Vicente, Vania A.
    Histoplasmosis is a systemic fungal disease that occurs worldwide, causing symptomatic infection mostly in immunocompromised hosts. Etiological agent is the dimorphic fungus, Histoplasma capsulatum, which occurs in soil contaminated with bird or bat droppings. Major limitation in recognition of H. capsulatum infections is the low awareness, since other diseases may have similar symptomatology. The molecular methods have gained importance because of unambiguous diagnostic ability and efficiency. The aim of this study was to develop and evaluate a padlock probe in view of rolling circle amplification (RCA) detection method which targets ITS (Internal Transcribed Spacer) rDNA of H. capsulatum enabling rapid and specific detection of the fungus in clinical samples. Two padlock probes were designed and one of these (HcPL2) allowed specific amplification of H. capsulatumDNA while no cross-reactivity was observed with fungi used as negative controls. This method proved to be effective for H. capsulatum specific identification and demonstrated to be faster than the traditional method of microbiological identification.
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    Molecular identification of Histoplasma capsulatum using rolling circle amplification
    (Wiley-Blackwell, 2016) Furuie, Jason L.; Sun, Jiufeng; do Nascimento, Mariana M. F.; Gomes, Renata Rodrigues; Waculicz-Andrade, Caroline E.; Sessegolo, Gisele C.; Rodrigues, Anderson Messias [UNIFESP]; Galvao-Dias, Maria Adelaide; Camargo, Zoilo Pires de [UNIFESP]; Queiroz-Telles, Flavio; Najafzadeh, Mohammad J.; de Hoog, Sybren G.; Vicente, Vania Aparecida; Universidade Federal de São Paulo (UNIFESP)
    Histoplasmosis is a systemic fungal disease that occurs worldwide, causing symptomatic infection mostly in immunocompromised hosts. Etiological agent is the dimorphic fungus, Histoplasma capsulatum, which occurs in soil contaminated with bird or bat droppings. Major limitation in recognition of H. capsulatum infections is the low awareness, since other diseases may have similar symptomatology. The molecular methods have gained importance because of unambiguous diagnostic ability and efficiency. The aim of this study was to develop and evaluate a padlock probe in view of rolling circle amplification (RCA) detection method which targets ITS (Internal Transcribed Spacer) rDNA of H. capsulatum enabling rapid and specific detection of the fungus in clinical samples. Two padlock probes were designed and one of these (HcPL2) allowed specific amplification of H. capsulatumDNA while no cross-reactivity was observed with fungi used as negative controls. This method proved to be effective for H. capsulatum specific identification and demonstrated to be faster than the traditional method of microbiological identification.