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

Publication date1983
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
  • Animals
  • Cell line
  • Cloning molecular
  • DNA genetics
  • Dopamine beta-hydroxylase genetics
  • Gene expression regulation
  • Nucleic acid hybridization
  • Peptide fragments analysis
  • RNA, messenger genetics
  • Rats
NotePMCID: PMC393777
Affiliation Not a UNIGE publication
Citation (ISO format)
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 of the United States of America, 1983, vol. 80, n° 8, p. 2161–2165.
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ISSN of the journal0027-8424
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