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Effect of growth hormone on fatty acid synthase gene expression in porcine adipose tissue cultures Genet. Mol. Biol.
José,Andrea A.F.B.V.; Alvares,Lúcia. E.; Delgado,Eduardo F.; Coutinho,Luiz L.; Lanna,Dante P.D..
We describe an efficient in vitro assay to test growth hormone effects on mRNA levels and fatty acid synthase (FAS, EC. 2.3.1.85) activity. Swine adipose tissue explants were long-term cultured in medium containing growth hormone and FAS mRNA levels and enzyme activity were measured. We quantified FAS transcripts by competitive reverse transcriptase PCR (RT-PCR) using total RNA from cultured adipose tissue explants and RT-PCR standard-curves were constructed using a cloned 307 bp segment of native FAS cDNA and a shorter fragment from which a 64 bp (competitor, 243 bp) internal sequence had been deleted. A known amount of competitor was added to each PCR as an internal control and µ-actin transcripts were also measured to correct for differences in total...
Tipo: Info:eu-repo/semantics/article Palavras-chave: MRNA quantification; Enzyme activity; Swine; Cultures of adipose tissue explants; Competitive RT-PCR.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572006000100032
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