RESUMO
Timely identification of biothreat organisms from large numbers of clinical or environmental samples in potential outbreak or attack scenario is critical for effective diagnosis and treatment. This study aims to evaluate the potential of resequencing arrays for this purpose. Albeit suboptimal, this report demonstrated that respiratory pathogen microarray version 1 can identify Bacillus anthracis, Francisella tularensis, Yersinia pestis and distinguish them from benign 'near neighbor' species in a single assay. Additionally, the sequence information can discriminate strains and possibly the sources of the strains. With further development, it is possible to use resequencing microarrays for biothreat surveillance.
Assuntos
Técnicas de Tipagem Bacteriana , Bioterrorismo/prevenção & controle , Francisella tularensis , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Análise de Sequência de DNA/métodos , Yersinia pestis , Antraz/diagnóstico , Antraz/microbiologia , Bacillus anthracis/classificação , Bacillus anthracis/genética , Bacillus anthracis/isolamento & purificação , Bacillus cereus/classificação , Bacillus cereus/genética , Bacillus cereus/isolamento & purificação , Método Duplo-Cego , Francisella/classificação , Francisella/genética , Francisella/isolamento & purificação , Francisella tularensis/classificação , Francisella tularensis/genética , Francisella tularensis/isolamento & purificação , Humanos , Peste/diagnóstico , Peste/microbiologia , Polimorfismo de Nucleotídeo Único , Especificidade da Espécie , Tularemia/diagnóstico , Tularemia/microbiologia , Yersinia/classificação , Yersinia/genética , Yersinia/isolamento & purificação , Yersinia pestis/classificação , Yersinia pestis/genética , Yersinia pestis/isolamento & purificaçãoRESUMO
Loblolly pine (Pinus taeda L.), the most widely planted tree species in the United States, is an important source of wood and wood fibers for a multitude of consumer products. Wood fibers are primarily composed of secondary cell walls, and cellulose, hemicelluloses and lignin are major components of wood. Fiber morphology and cell wall composition are important determinants of wood properties. We used comparative genomics to identify putative genes for cellulose and hemicellulose synthesis in loblolly pine that are homologous to genes implicated in cell wall synthesis in angiosperms. Sequences encoding putative secondary cell wall cellulose synthase genes, cellulose synthase-like genes, a membrane-bound endoglucanase gene, a sucrose synthase gene, a UDP-glucose pyrophosphorylase gene and GDP-mannose pyrophosphorylase genes were identified in expressed sequence tag (EST) collections from loblolly pine. Full-length coding sequences were obtained from cDNA clones isolated from a library constructed from developing xylem. Phylogenetic relationships between the genes from loblolly pine and angiosperm taxa were examined and transcriptional profiling in vascular tissues was conducted by real-time quantitative, reverse transcriptase-polymerase chain reaction. The putative cell wall synthesis genes were expressed at high levels in vascular tissues and a subset was differentially regulated in xylem and phloem tissues. Inferred phylogenetic relationships and expression patterns for the genes from loblolly pine were consistent with roles in synthesis of complex carbohydrates of the cell wall. These studies suggest functional conservation of homologous wood formation genes in gymnosperm and angiosperm taxa.
