title: Molecular cloning and domain structure of chicken pyruvate carboxylase.
abstract: Pyruvate carboxylase (PC) [EC 6.4.1.1] is a biotin-dependent carboxylase that catalyses the conversion of pyruvate to oxaloacetate. Here we have determined the complete nucleotide sequence encoding chicken PC (cPC) by screening a liver cDNA library, by RT-PCR of poly(A)(+) RNA, and by PCR of genomic DNA. The full-length transcript contains an open reading frame of 3537 nucleotides, including the stop codon, encoding a polypeptide of 1178 amino acids with M(r) of 127,262. The amino acid sequence of cPC shows approximately 77% identity to mammalian PC. Limited proteolysis of pure cPC with chymotrypsin yields a major stable 75 kDa C-terminal peptide, including the biotinyl domain and a minor, unstable 39 kDa N-terminal peptide. Northern analysis of poly(A)(+) RNA isolated from chicken liver has shown that cPC's mRNA is approximately 5 kb in length, including a very long 3'-untranslated region.
authors.affiliation: Department of Molecular Biosciences, Adelaide University, Adelaide, SA 5005, Australia.
authors.forename: Sarawut
authors.initials: S
authors.lastname: Jitrapakdee
authors.forename: Mark G
authors.initials: MG
authors.lastname: Nezic
authors.forename: A Ian
authors.initials: AI
authors.lastname: Cassady
authors.forename: Yeesim
authors.initials: Y
authors.lastname: Khew-Goodall
authors.forename: John C
authors.initials: JC
authors.lastname: Wallace
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002645:Chickens; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008969:Molecular Sequence Data; D011766:Pyruvate Carboxylase / Q000737:chemistry / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D011766:Pyruvate Carboxylase