Molecular cloning and nucleotide sequence analysis of complementary DNA for bullfrog prolactin

N. Takahashi, K. Yoshihama, S. Kikuyama, K. Yamamoto, K. Wakabayashi, Y. Kato

    Research output: Contribution to journalArticle

    32 Citations (Scopus)

    Abstract

    A prolactin cDNA was clonsed from a cDNA expression library constructed from total RNA of bullfrog (Rana catesbeiana) adenohypophyses by immunoscreening with antiserum against bullfrog prolactin. The cDNA clone thus obtained contained a 249 bp insert. Using this clone as a probe, plaque hybridizations were performed and two additional clones obtained. These clones had a polyadenylation site different from that of the first obtained clone, suggesting that the 3'-untranslated sequence was heterogeneous in length. The longest clone contained 830 bp, which encoded part of the signal peptide and the entire sequence of mature prolactin. The deduced amino acid sequence was in good accord with that determined by direct protein sequencing of purified bullfrog prolactin. The length of the bullfrog prolactin mRNA was estimated by Northern blot analysis to be about 1.0 kb. Homologies of prolactin nucleotide and amino acid sequences between bullfrog and other vertebrates were 64 and 65% for man, 66 and 68% for pig, 61 and 52% for rat, 69 and 74% for chicken, and 50 and 35% for salmon respectively. Highly conserved regions reported for mammalian prolactins also existed in bullfrog prolactin. Homologies of nucleotide and amino acid sequences between prolactin and GH of bullfrog origin were 49 and 25% respectively. Using the cDNA, the content of prolactin mRNA in the pituitary glands of metamorphosing tadpoles was measured. Prolactin mRNA levels rose at the mid-climax stage, suggesting that the increase in plasma and pituitary prolactin levels known to occur at the climax stage accompanies the increase in prolactin synthesis.

    Original languageEnglish
    Pages (from-to)281-287
    Number of pages7
    JournalJournal of Molecular Endocrinology
    Volume5
    Issue number3
    Publication statusPublished - 1990

    Fingerprint

    Rana catesbeiana
    Molecular Cloning
    Prolactin
    Sequence Analysis
    Complementary DNA
    Clone Cells
    Messenger RNA
    Amino Acid Sequence
    Nucleotides
    Amino Acid Sequence Homology
    Polyadenylation
    Anterior Pituitary Gland
    Salmon
    Protein Sequence Analysis
    Pituitary Gland
    Protein Sorting Signals
    Gene Library
    Northern Blotting
    Larva
    Vertebrates

    ASJC Scopus subject areas

    • Endocrinology

    Cite this

    Takahashi, N., Yoshihama, K., Kikuyama, S., Yamamoto, K., Wakabayashi, K., & Kato, Y. (1990). Molecular cloning and nucleotide sequence analysis of complementary DNA for bullfrog prolactin. Journal of Molecular Endocrinology, 5(3), 281-287.

    Molecular cloning and nucleotide sequence analysis of complementary DNA for bullfrog prolactin. / Takahashi, N.; Yoshihama, K.; Kikuyama, S.; Yamamoto, K.; Wakabayashi, K.; Kato, Y.

    In: Journal of Molecular Endocrinology, Vol. 5, No. 3, 1990, p. 281-287.

    Research output: Contribution to journalArticle

    Takahashi, N, Yoshihama, K, Kikuyama, S, Yamamoto, K, Wakabayashi, K & Kato, Y 1990, 'Molecular cloning and nucleotide sequence analysis of complementary DNA for bullfrog prolactin', Journal of Molecular Endocrinology, vol. 5, no. 3, pp. 281-287.
    Takahashi N, Yoshihama K, Kikuyama S, Yamamoto K, Wakabayashi K, Kato Y. Molecular cloning and nucleotide sequence analysis of complementary DNA for bullfrog prolactin. Journal of Molecular Endocrinology. 1990;5(3):281-287.
    Takahashi, N. ; Yoshihama, K. ; Kikuyama, S. ; Yamamoto, K. ; Wakabayashi, K. ; Kato, Y. / Molecular cloning and nucleotide sequence analysis of complementary DNA for bullfrog prolactin. In: Journal of Molecular Endocrinology. 1990 ; Vol. 5, No. 3. pp. 281-287.
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    abstract = "A prolactin cDNA was clonsed from a cDNA expression library constructed from total RNA of bullfrog (Rana catesbeiana) adenohypophyses by immunoscreening with antiserum against bullfrog prolactin. The cDNA clone thus obtained contained a 249 bp insert. Using this clone as a probe, plaque hybridizations were performed and two additional clones obtained. These clones had a polyadenylation site different from that of the first obtained clone, suggesting that the 3'-untranslated sequence was heterogeneous in length. The longest clone contained 830 bp, which encoded part of the signal peptide and the entire sequence of mature prolactin. The deduced amino acid sequence was in good accord with that determined by direct protein sequencing of purified bullfrog prolactin. The length of the bullfrog prolactin mRNA was estimated by Northern blot analysis to be about 1.0 kb. Homologies of prolactin nucleotide and amino acid sequences between bullfrog and other vertebrates were 64 and 65{\%} for man, 66 and 68{\%} for pig, 61 and 52{\%} for rat, 69 and 74{\%} for chicken, and 50 and 35{\%} for salmon respectively. Highly conserved regions reported for mammalian prolactins also existed in bullfrog prolactin. Homologies of nucleotide and amino acid sequences between prolactin and GH of bullfrog origin were 49 and 25{\%} respectively. Using the cDNA, the content of prolactin mRNA in the pituitary glands of metamorphosing tadpoles was measured. Prolactin mRNA levels rose at the mid-climax stage, suggesting that the increase in plasma and pituitary prolactin levels known to occur at the climax stage accompanies the increase in prolactin synthesis.",
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