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Title

Genes for catecholamine biosynthesis : cloning by expression and identification of the cDNA for rat dopamine beta-hydroxylase

Authors
O'Malley, K. L.
Raese, J.
Barchas, J. D.
Kedes, L.
Published in Proceedings of the National Academy of Sciences. 1983, vol. 80, no. 8, p. 2161-2165
Abstract mRNA for dopamine beta-hydroxylase [3,4-dihydroxyphenylethylamine, ascorbate:oxygen oxidoreductase (beta-hydroxylating), EC 1.14.17.1] has been partially purified from poly(A)+ mRNA isolated from a rat pheochromocytoma cell line. Shared antigenic determinants between tyrosine hydroxylase and dopamine beta-hydroxylase allowed us to obtain enriched fractions of dopamine beta-hydroxylase mRNA by immunoprecipitating translated mRNA products with tyrosine hydroxylase antisera. The enriched dopamine beta-hydroxylase mRNA was used to synthesize the corresponding cDNAs, which were then cloned in the Pst I site of pBR322. Recombinant colonies were characterized by an in situ colony immunoassay and hybrid-selected translation. In vitro translation of the mRNA selected from one recombinant clone produced a protein of 75,000 daltons that comigrated with authentic dopamine beta-hydroxylase. Partial proteolysis of both authentic dopamine beta-hydroxylase and the protein encoded by the recombinant clone produced identical peptide patterns.
Keywords AnimalsCell lineCloning molecularDNA geneticsDopamine beta-hydroxylase geneticsGene expression regulationNucleic acid hybridizationPeptide fragments analysisRNA, messenger geneticsRats
Identifiers
PMID: 6572968
Note PMCID: PMC393777
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O'MALLEY, K. L. et al. Genes for catecholamine biosynthesis : cloning by expression and identification of the cDNA for rat dopamine beta-hydroxylase. In: Proceedings of the National Academy of Sciences, 1983, vol. 80, n° 8, p. 2161-2165. https://archive-ouverte.unige.ch/unige:85378

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Deposited on : 2016-07-20

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