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Obtención de una sonda para cuantificar el mARN del factor activador de la transcripción STAT5 en rata

1998; National Institute of Health; Volume: 18; Issue: 3 Linguagem: Inglês

10.7705/biomedica.v18i3.989

2590-7379

Blanca L. Ortíz, Myriam Sánchez-Gómez, Gunnar Norstedt,

Protein Tyrosine Phosphatases

The strategy for cloning a PCR-amplified STAT5 fragment into an expression vector is described. By optimising the magnesium level and the annealing temperature in the PCR reaction, target fragment amplification was achieved, using mouse STAT5b cDNA as a template. The PCR product was cloned and probe identity was confirmed by restriction analysis and sequencing. This probe was used in a solution hybridisation-Rnase protection assay to quantify STAT5 mRNA in female rats' livers and thymus lymphocytes. A higher STAT5 mRNA expression was found in liver.