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Mar Biotechnol (NY) ; 5(6): 545-54, 2003.
Artículo en Inglés | MEDLINE | ID: mdl-14508654

RESUMEN

Commercially available human complementary DNA microarrays were used to compare differential expression in the livers of Atlantic salmon ( Salmo salar) infected with Aeromonas salmonicida and of healthy fish. Complementary DNA probes were prepared from total RNA isolated from livers of control salmon and infected salmon by reverse transcription in the presence of (33)P-dCTP and independently hybridized to human GENE-FILTERS GF211 microarrays. Of the 4131 known genes on the microarray, 241 spots gave clearly detectable signals using labeled RNA from the control salmon liver. Of these, 4 spots were consistently found to have a greater than 2-fold increase in infected salmon compared with controls when using the same pair of filters to generate hybridization data from triplicates. These up-regulated genes were ADP/ATP translocase (AAT2), Na(+)/K(+) ATPase, acyloxyacyl hydrolase (AOAH), and platelet-derived growth factor (PDFG-A). A BlastN search revealed an AAT2 homolog from Atlantic salmon, and a reverse transcriptase polymerase chain reaction assay using primers based on this sequence confirmed its up-regulation (approx. 1.8-fold) during early infection. This work demonstrates the feasibility of using human microarrays to facilitate the discovery of differentially expressed genes in Atlantic salmon, for which no homologous microarrays are available.


Asunto(s)
Aeromonas , Enfermedades de los Peces/microbiología , Infecciones por Bacterias Gramnegativas/veterinaria , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Salmo salar/genética , Regulación hacia Arriba , Secuencia de Aminoácidos , Animales , Hidrolasas de Éster Carboxílico/genética , Cartilla de ADN , Bases de Datos Genéticas , Enfermedades de los Peces/genética , Perfilación de la Expresión Génica , Humanos , Hígado/química , Translocasas Mitocondriales de ADP y ATP/genética , Datos de Secuencia Molecular , Factor de Crecimiento Derivado de Plaquetas/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , ATPasa Intercambiadora de Sodio-Potasio/genética
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