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Cloning a fragment from the telomere of the long arm of human chromosome 9 in a YAC vector

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Abstract

The construction of a yeast artificial chromosome containing a human DNA insert is reported. This molecule of about 200 kb behaves as a native yeast chromosome since it has a very high mitotic stability and is present in the yeast transformant clone at a copy number similar to that of the resident chromosomes. Hybridization with the TTAGGG sequence demonstrates that this chromosome contains human telomeric sequences. In situ hybridization of the biotin-labelled artificial chromosome to metaphase human chromosomes shows that the insert occupies a telomeric position on the long arm of chromosome 9. Since the fragment was cloned as an EcoRI insert and not as a telomere, it is situated medially to the telomeric sequences and harbours telomere-associated sequences, that have been shown to contain the TTAGGG sequence. The fragment represents the end of the genetic map of chromosome 9 and thus can be used to characterize the sequence and the structure of the chromosomal region that runs from the end of the chromosome to the first gene.

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References

  • Allshire RC, Dempster M, Hastie ND (1989) Human telomeres contain at least three types of G-rich repeat distributed nonrandomly. Nucleic Acids Res 17:4611–4627

    Google Scholar 

  • Blackburn EH, Szostak JW (1984) The molecular structures of centromeres and telomeres. Annu Rev Biochem 53:163–194

    Google Scholar 

  • Brown WRA (1989) Molecular cloning of human telomeres in yeast. Nature 38:774–776

    Google Scholar 

  • Burke DT, Carle GF, Olson MV (1987) Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vector. Science 236:806–811

    Google Scholar 

  • Carle CF, Olson MV (1985) An electrophoretic karyotype of yeast. Proc Natl Acad Sci USA 82:3756–3760

    Google Scholar 

  • Chan CSM, Tye BK (1983) Organization of DNA sequences and replication origin at yeast telomeres. Cell 33:563–573

    Google Scholar 

  • Cheng J-F, Smith LC, Cantor CR (1989) Isolation and characterization of a human telomere. Nucleic Acids Res 17:6109–6125

    Google Scholar 

  • Cross SH, Allshire RC, Mckey SJ, McGill NJ, Cook HJ (1989) Cloning of human telomeres by complementation in yeast. Nature 338:771–774

    Google Scholar 

  • Cryer DR, Eccleshall R, Marmur J (1975) Isolation of yeast DNA. Methods Cell Biol 12:39–44

    Google Scholar 

  • Greider CW, Blackburn EH (1987) The telomere terminal transferase of Tetrahymena is a ribonucleoprotein enzyme with two distinct primer specificity components. Cell 51:887–898

    Google Scholar 

  • Greider CW, Blackburn EH (1989) A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis. Nature 337:331–337

    Google Scholar 

  • Guerrini AM, Ascenzioni F, Tribioli C, Donini P (1985) Transformation ofSaccharomyces cerevisiae andSchizosaccharomyces pombe with linear plasmids containing the 2 µ sequences. EMBO J 4:1569–1573

    Google Scholar 

  • Henderson E, Hardin CC, Walk SK, Tinoco I, Blackburn EH (1987) Telomeric DNA oligonucleotides form novel intramolecular structures containing guanine-guanine base pairs. Cell 51:899–908

    Google Scholar 

  • Klobutcher LT, Swanton MT, Donini P, Prescott DM (1981) All gene-sized DNA molecules in four species of hypotrichs have the same terminal sequence and an unusual 3′ terminus. Proc Natl Acad Sci USA 78:3015–3019

    Google Scholar 

  • Lyamichev VI, Mirkin SM, Danilevskaya ON, Voloshin ON, Valatskaya SV, Dobrynin VN, Filippov SA, Frank-Kamenetskii MD (1989) An unusual DNA structure detected in a telomeric sequence under superhelical stress and at low pH. Nature 339:634–637

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory, NY, p 122

    Google Scholar 

  • Moyzis RK, Buckingham JM, Cram LS, Dani M, Deaven LL, Jones MD, Myne J, Ratliff RL, Wu J-R (1988) A highly conserved repetitive DNA sequence (TTAGGG)n present at the telomeres of human chromosomes. Proc Natl Acad Sci USA 85:6622–6626

    Google Scholar 

  • Murray AW, Schultes NP, Szostak JW (1986) Chromosome length controls mitotic segregation in yeast. Cell 43:529–536

    Google Scholar 

  • Ponzi M, Pace T, Dore E, Frontali C (1987) Identification of telomeric DNA sequence in P. berghei. EMBO J 4:2991–2995

    Google Scholar 

  • Szostak JW, Blackburn EH (1982) Cloning yeast telomeres on linear plasmid vectors. Cell 29:245–255

    Google Scholar 

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Guerrini, A.M., Ascenzioni, F., Pisani, G. et al. Cloning a fragment from the telomere of the long arm of human chromosome 9 in a YAC vector. Chromosoma 99, 138–142 (1990). https://doi.org/10.1007/BF01735330

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  • DOI: https://doi.org/10.1007/BF01735330

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