Abstract
Telomeres are specialized structures that form the ends of eukaryotic chromosomes and that are required to fulfill a number of varied functions, see Biessman and Mason for recent review (1). First, they must protect the chromosome ends from degradation and from fusion and recombination with other chromosome ends or internal DNA. They must also provide a solution to the problem of chromosome end replication, because all known polymerases require a primer and synthesize DNA from 5′-3′, therefore, the 3′ ends of linear DNA pose a problem to the normal replication machinery. They can influence gene expression by position effect variagation and finally, telomeres may play a role in establishing the architecture of the interphase and/or the meiotic prophase nucleus. Isolation of telomeric sequences is therefore of interest with respect to unraveling telomere structure and the mechanisms underlying telomeric function. In addition, the isolation and characterization of all human telomeres is a necessary part of the human genome project in order to generate genetic and physical markers at the ends of chromosomes thereby providing the boundaries of genetic and physical maps.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Biessmann, H. and Mason, J. M. (1992) Genetics and molecular biology of telomeres. Adv. Genet. 30, 185–249.
Zakian, V. A. (1989) Structure and function of telomeres. Annu. Rev. Genet. 23, 579–604.
Meyne, J., Ratcliffe, R. L., and Moyzis, R. K. (1989) Conservation of the human telomere sequence (TTAGGG)n among vertebrates. Proc. Natl. Acad. Sci. USA 86, 7049–7053.
Szostak, J. W. and Blackburn, E. H. (1982) Cloning yeast telomeres on a linear plasmid. Cell 29, 245–255.
Burke, D. T., Carle, G. F., and Olson, M. V. (1987) Cloning of large DNA segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science 236, 806–812.
Moyzis, R. K., Buckingham, J. M., Cram, L. S., Dam, M., Deaven, L. L., Jones, M. D., et al. (1988) A highly conserved repetitive DNA sequence, (TTAGGG)n, present at the telomeres of human chromosomes. Proc. Natl. Acad. Sci. USA 85, 6622–6626.
Bates, G. P., MacDonald, M. E., Baxendale, S., Sedlacek, Z., Youngman, S., Romano, D., et al. (1990) A yeast artificial chromosome telomere clone spanning a possible location of the Huntington disease gene. Am. J. Hum. Genet. 46, 762–775.
Brown, W. R. A. (1989) Molecular cloning of human telomeres in yeast. Nature 338, 774–776.
Cheng, J.-F., Smith, C. L., and Cantor, C. R. (1989) Isolation and characterisation of a human telomere. Nucleic Acids Res. 17, 6109–6127.
Cross, S. H., Allshire, R. C., McKay, S. J., McGill, N. I., and Cooke, H. J. (1989) Cloning of human telomeres by complementation in yeast. Nature 338, 771–774.
Riethman, H. C., Moyzis, R. K., Meyne, J., Burke, D. T., and Olsen, M. V. (1989) Cloning human telomeric DNA fragments into Saccharomyces cerevisiae using a yeast-artificial-chromosome vector. Proc. Natl. Acad. Sci. USA 86, 6240–6244.
Youngman, S., Bates, G. P., Williams, S., McClatchey, A. I., Baxendale, S., Sedlacek, Z., et al. (1992) The telomeric 60 kb of chromosome arm 4p is homologous to telomeric regions on 13p, 15p, 21p, and 22p. Genomics 14, 350–356.
Hieter, P., Mann, C., Snyder, M., and Davis, R. W. (1985) Mitotic stability of yeast chromosomes: a colony color assay that measures nondisjunction and chromosome loss. Cell 40, 381–392.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Herrmann, B. G., Barlow, D. P. and Lehrach, H. (1987) An inverted duplication of more than 650 Kbp in mouse chromosome 17 mediates unequal but homologous recombination between chromosomes heterozygous for a large inversion. Cell 48, 813–825.
Herrmann, B. G. and Frischauf, A.-M. (1987) Isolation of genomic DNA, Academic, New York.
Burgers, P. M. J. and Percival, K. J. (1987) Transformation of yeast spheroplasts without cell fusion. Anal. Biochem. 163, 391–397.
Brown, W. R. A., MacKinnon, P. J., Villasante, A., Spurr, N., Buckle, V. J., and Dobson, M. J. (1990) Structure and polymorphism of human telomere-associated DNA. Cell 63, 119–132.
de Lange, T., Shiue, L., Myers, R. M., Cox, D. R., Naylor, S. L., Killery, A. M., and Varmus, H. E. (1990) Structure and variability of human chromosome ends. Mol. Cell Biol. 10, 518–527.
Larin, Z., Monaco, A. P., and Lehrach, H. (1991) Yeast artificial chromosome libraries containing large inserts from mouse and human DNA. Proc. Natl. Acad. Sci. USA 88, 4123–4127.
Smith, D. R., Smyth, A. P., and Moir, D. T. (1990) Amplification of large artificial chromosomes. Proc. Natl. Acad. Sci. USA 87, 8242–8246.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1996 Humana Press Inc.
About this protocol
Cite this protocol
Bates, G. (1996). Cloning of Human Telomeres in Saccharomyces cerevisiae . In: Markie, D. (eds) YAC Protocols. Methods in Molecular Biology™, vol 54. Humana Press. https://doi.org/10.1385/0-89603-313-9:49
Download citation
DOI: https://doi.org/10.1385/0-89603-313-9:49
Publisher Name: Humana Press
Print ISBN: 978-0-89603-313-9
Online ISBN: 978-1-59259-541-9
eBook Packages: Springer Protocols