Obtaining a specific probe to evaluate STAT5b gene transcription
Keywords:
RT-PCR, GH, STAT5b, solution hybridization assay
Abstract
The strategy for cloning a 306 bp rat STAT5b fragment, amplified by RT-PCR, into an expression vector is described.The main difference between STAT5a and STAT5b lies within the 3' region of their respective cDNAs, therefore a fragment in this region was amplified. Cloning was achieved by using the new system TOPO-TA recommended, for products difficult to be cloned.The fragment was subcloned into pBlueScript Il SK vector and characterized by sequencing and restriction analysis. Subcloning was done in order to obtain sense and antisense RNA probes by in vitro transcription. This specific probe has been used to quantify STAT5b mRNA levels in liver from male rats, by a soluiiori hybridization RNAase protection assay.Downloads
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How to Cite
1.
Díaz LE, Ortiz BL, Sánchez de Gómez M. Obtaining a specific probe to evaluate STAT5b gene transcription. biomedica [Internet]. 2001 Jun. 1 [cited 2024 May 18];21(2):142-9. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/1102
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Published
2001-06-01
Issue
Section
Original articles
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