Isolation and characterization of a cDNA encoding a pea ornithine transcarbamoylase (argF) and comparison with other transcarbamoylases.
We used a PCR-based library screening method to isolate a 1.4 kb pea leaf cDNA encoding ornithine transcarbamoylase (OTCase). The cDNA contains a single major ORF of 375 amino acids whose deduced sequence exhibits a high degree of homology with other OTCases. The predicted molecular mass of 41361 Da for this protein is approximately the 40 kDa size of the polypeptide that is immunoprecipitated with OTCase antibody after in vitro translation of pea leaf mRNA. In vivo, OTCase occurs as a trimer of identical 36.5 kDa polypeptides, suggesting that this enzyme is synthesized as a cytosolic precursor protein. Southern blot analysis indicates that multiple OTCase genes occur in pea. An abundant 1.4 kb transcript is seen in northern blots of total RNA isolated from the leaves and roots of light- and dark-grown pea seedlings.
Department of Biological Sciences, Goucher College, Baltimore, MD 21204-2794 USA.
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Williamson
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Lake
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D000595:Amino Acid Sequence; D000818:Animals; D003429:Cross Reactions; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics*; D015723:Gene Library; D018745:Genome, Plant; D051379:Mice; D008969:Molecular Sequence Data; D009954:Ornithine Carbamoyltransferase / Q000235:genetics*; D018532:Pisum sativum / Q000201:enzymology / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D011498:Protein Precursors / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D017931:DNA Primers; D018076:DNA, Complementary; D011498:Protein Precursors; D012333:RNA, Messenger; D018749:RNA, Plant; D009954:Ornithine Carbamoyltransferase