PROIMI   05436
PLANTA PILOTO DE PROCESOS INDUSTRIALES MICROBIOLOGICOS
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Genetic expression by real time PCR of bgl-A, bgl, CspA, gyrB and 16S ADNr genes from Shewanella sp G5
Autor/es:
CRISTÓBAL, H. A.; POMA, H. R.; RAJAL, V. B.; ABATE, C. M.
Lugar:
Villa Carlos Paz, Córdoba
Reunión:
Congreso; XLIV Reunión Anual de la Sociedad Argentina de Investigaciones Bioquímicas; 2008
Institución organizadora:
Sociedad Argentina de Investigaciones Bioquímicas
Resumen:
Shewanella sp. G5 is a psychrotolerant Gram negative bacteria ¥â-glucosidases (EC 3.2.1.21) producers. This marine bacterium was isolate from the intestinal content of Munida subrrugosa in the Beagle Channel, Tierra del Fuego (Argentina); and grows between 4 and 37 ¨¬C using cellobiose as carbon source. ¥â-glucosidases are a heterogeneous group of enzymes with a broad substrate specificity range over different aryl- and alkyl-¥â-D-glucosides. The gene (bglA and bgl) that encoding of two ¥â-glucosidases were characterized molecularly in previous studies. The primers design, RNA extraction, synthesis of cDNA and real time PCR assays using SYBRGreen (Invitrogen) were performed. The parameter assays to each condition were: temperature (10 and 30 ¨¬C), carbon source (cellobiose and glucose) and culture media. bglA, bgl, CspA, gyrB and ADNr 16S genes were quantified and expressed as relative genetic expression from of each assays using 2-¥Ä¥ÄCt method.  Positive results were obtained for all genes during the optimization of the real time PCR. The amplification was verified by standard and dissociation curves and amplot analysis (reporter of Ct). Normalization assays using 2-¥Ä¥ÄCt method were carried out with the housekeeping genes (gyrB and ADNr 16S) also.  The best values of relative genetic expression index for  bglA and bgl were 27,67 and 30,17 respectively; and were obtained at the following assayed conditions: Brunner medium, 30 ¨¬C and in presence of cellobiose.