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Proc Natl Acad Sci U S A. 1985 May; 82(9): 2613–2617.
PMCID: PMC397614

Antifreeze protein genes are tandemly linked and clustered in the genome of the winter flounder.


We have used genomic Southern blots and restriction maps of genomic clones to examine the organization of the antifreeze protein multigene family in the winter flounder. The majority of the approximately equal to 40 antifreeze protein (AFP) genes in this fish are present in 7- to 8-kilobase-pair (kbp) elements of DNA, which are iterated as tandem direct repeats. Each repeat contains a single antifreeze protein gene that is 1 kbp long, and all of these genes have the same transcriptional orientation. Although the repeated elements are highly homologous, they do show some restriction site and restriction length polymorphisms. When flounder genomic DNA is digested with restriction endonucleases that do not cut within the repeats, most of the antifreeze protein genes reside in fragments that are at least 40 kbp long, representing clusters of five or more repeats in tandem. After genomic DNA is digested with Xba I or Xho I, these genes are present in fragments of exceptionally high molecular weight, suggesting that the clusters themselves are grouped together in the genome. The AFP gene locus may have evolved by gene amplification as recently as 10(6) years ago in response to the onset of the Cenozoic ice age.

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  • DeVries AL. Antifreeze peptides and glycopeptides in cold-water fishes. Annu Rev Physiol. 1983;45:245–260. [PubMed]
  • Duman JG, de Vries AL. Isolation, characterization, and physical properties of protein antifreezes from the winter flounder, Pseudopleuronectes americanus. Comp Biochem Physiol B. 1976;54(3):375–380. [PubMed]
  • Hew CL, Yip C. The synthesis of freezing-point-depressing protein of the winter flounder Pseudopleuronectus americanus in Xenopus laevis oocytes. Biochem Biophys Res Commun. 1976 Aug 9;71(3):845–850. [PubMed]
  • Fletcher GL. Circannual cycles of blood plasma freezing point and Na+ and Ci- concentrations in Newfoundland winter flounder (Pseudopleuronectes americanus): correlation with water temperature and photoperiod. Can J Zool. 1977 May;55(5):789–795. [PubMed]
  • Devries AL, Lin Y. Structure of a peptide antifreeze and mechanism of adsorption to ice. Biochim Biophys Acta. 1977 Dec 20;495(2):388–392. [PubMed]
  • Davies PL, Roach AH, Hew CL. DNA sequence coding for an antifreeze protein precursor from winter flounder. Proc Natl Acad Sci U S A. 1982 Jan;79(2):335–339. [PMC free article] [PubMed]
  • Slaughter D, Hew CL. Radioimmunoassay for the antifreeze polypeptides of the winter flounder: seasonal profile and immunological cross-reactivity with other fish antifreezes. Can J Biochem. 1982 Aug;60(8):824–829. [PubMed]
  • Pickett MH, Hew CL, Davies PL. Seasonal variation in the level of antifreeze protein mRNA from the winter flounder. Biochim Biophys Acta. 1983 Jan 20;739(1):97–104. [PubMed]
  • Fourney RM, Fletcher GL, Hew CL. Accumulation of winter flounder antifreeze messenger RNA after hypophysectomy. Gen Comp Endocrinol. 1984 Jun;54(3):392–401. [PubMed]
  • Davies PL, Hough C, Scott GK, Ng N, White BN, Hew CL. Antifreeze protein genes of the winter flounder. J Biol Chem. 1984 Jul 25;259(14):9241–9247. [PubMed]
  • Schaffner W, Gross K, Telford J, Birnstiel M. Molecular analysis of the histone gene cluster of psammechinus miliaris: II. The arrangement of the five histone-coding and spacer sequences. Cell. 1976 Aug;8(4):471–478. [PubMed]
  • Long EO, Dawid IB. Repeated genes in eukaryotes. Annu Rev Biochem. 1980;49:727–764. [PubMed]
  • Thomashow LS, Milhausen M, Rutter WJ, Agabian N. Tubulin genes are tandemly linked and clustered in the genome of trypanosoma brucei. Cell. 1983 Jan;32(1):35–43. [PubMed]
  • Blin N, Stafford DW. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. [PMC free article] [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Pickett M, Scott G, Davies P, Wang N, Joshi S, Hew C. Sequence of an antifreeze protein precursor. Eur J Biochem. 1984 Aug 15;143(1):35–38. [PubMed]
  • Benton WD, Davis RW. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. [PubMed]
  • Callan HG. The organization of genetic units in chromosomes. J Cell Sci. 1967 Mar;2(1):1–7. [PubMed]
  • Tartof KD. Unequal mitotic sister chromatid exchange and disproportionate replication as mechanisms regulating ribosomal RNA gene redundancy. Cold Spring Harb Symp Quant Biol. 1974;38:491–500. [PubMed]
  • Szostak JW, Wu R. Unequal crossing over in the ribosomal DNA of Saccharomyces cerevisiae. Nature. 1980 Apr 3;284(5755):426–430. [PubMed]
  • Petes TD. Unequal meiotic recombination within tandem arrays of yeast ribosomal DNA genes. Cell. 1980 Mar;19(3):765–774. [PubMed]
  • Roberts JM, Axel R. Gene amplification and gene correction in somatic cells. Cell. 1982 May;29(1):109–119. [PubMed]
  • Klein HL, Petes TD. Intrachromosomal gene conversion in yeast. Nature. 1981 Jan 15;289(5794):144–148. [PubMed]
  • Jackson JA, Fink GR. Gene conversion between duplicated genetic elements in yeast. Nature. 1981 Jul 23;292(5821):306–311. [PubMed]
  • Ericson DB, Wollin G. Pleistocene climates and chronology in deep-sea sediments. Science. 1968 Dec 13;162(3859):1227–1234. [PubMed]
  • Slaughter D, Fletcher GL, Ananthanarayanan VS, Hew CL. Antifreeze proteins from the sea raven, Hemitripterus americanus. Further evidence for diversity among fish polypeptide antifreezes. J Biol Chem. 1981 Feb 25;256(4):2022–2026. [PubMed]
  • Hew CL, Fletcher GL, Ananthanarayanan VS. Antifreeze proteins from the shorthorn sculpin, Myoxocephalus scorpius: isolation and characterization. Can J Biochem. 1980 May;58(5):377–383. [PubMed]

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