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. 2000 Aug;67(2):320-32.
doi: 10.1086/302998. Epub 2000 Jun 22.

An optimized set of human telomere clones for studying telomere integrity and architecture

S J Knight  1 , C M Lese, K S Precht, J Kuc, Y Ning, S Lucas, R Regan, M Brenan, A Nicod, N M Lawrie, D L Cardy, H Nguyen, T J Hudson, H C Riethman, D H Ledbetter, J Flint
Affiliations

An optimized set of human telomere clones for studying telomere integrity and architecture

S J Knight et al. Am J Hum Genet. 2000 Aug.

Abstract

Telomere-specific clones are a valuable resource for the characterization of chromosomal rearrangements. We previously reported a first-generation set of human telomere probes consisting of 34 genomic clones, which were a known distance from the end of the chromosome ( approximately 300 kb), and 7 clones corresponding to the most distal markers on the integrated genetic/physical map (1p, 5p, 6p, 9p, 12p, 15q, and 20q). Subsequently, this resource has been optimized and completed: the size of the genomic clones has been expanded to a target size of 100-200 kb, which is optimal for use in genome-scanning methodologies, and additional probes for the remaining seven telomeres have been identified. For each clone we give an associated mapped sequence-tagged site and provide distances from the telomere estimated using a combination of fiberFISH, interphase FISH, sequence analysis, and radiation-hybrid mapping. This updated set of telomeric clones is an invaluable resource for clinical diagnosis and represents an important contribution to genetic and physical mapping efforts aimed at telomeric regions.

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Figures

Figure 1
Figure 1
Comparison of FISH results obtained with a cosmid clone compared to a BAC clone for the 19p and 10q telomeres. All probes are labeled with digoxigenin and detected with antidigoxigenin rhodamine (red). A, Hybridization of cosmid clone, cF20643 (left), and BAC clone, RG129F16 (right), corresponding to the 19p telomere. B, Hybridization of cosmid clone, c2136c3 (left), and BAC clone, GS261B16 (right), corresponding to the 10q telomere. In both A and B, notice the difference in signal intensity between the two telomere clones; the BAC clone shows a larger signal compared to that of the cosmid clone.
Figure 2
Figure 2
Three-color interphase FISH ordering at the human 1p telomere. Interphase cells from a chromosome 1 monochromosomal hybrid cell line were hybridized with three probes from the telomeric region of chromosome 1p. GS-232-B23 is labeled with biotin and detected with avidin-Cy5 (purple), PAC GS-63-M14 is labeled with Spectrum Orange (red) and BAC GS-145-I23 is labeled with digoxigenin and detected with anti-digoxigenin FITC (green). The order of these clones was determined to be tel-GS145I23-GS63M14-GS232B23-cen.
Figure 3
Figure 3
Diagram showing the derivation of fiberFISH distances. The fiberFISH distance in kb (D) between the test clone signal and the telomere (TTAGGGital)n signal is given by D=Z(X/Y), where Z is the known size of the test clone in kb, Y is the length of the test-clone signal in mm and X is the distance between the test-clone signal and the telomere signal in mm.
Figure 4
Figure 4
FiberFISH results showing telomere-specific probe signals in green and (TTAGGG)n probe signals in red. A, Control hybridization showing 4p cosmid (B31: ∼33 kb insert) ∼80 kb from the (TTAGGG)n repeat sequence. B, control hybridization showing 16p cosmid (cGG4: ∼36 kb insert) ∼100 kb from the (TTAGGG)n repeat. C, test hybridization showing 7p PAC (164-D18) ∼130 kb from the (TTAGGG)n repeat.
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References

Electronic-Database Information

    1. Bacpac Web site, http://www.chori.org/bacpac/home.htm
    1. Comprehensive genetic map, http://www.marshmed.org/genetics/ (for distal marker identification)
    1. GenBank, http://www.ncbi.nlm.gov/Genbank/GenbankSearch.html (for primers and sequence information)
    1. Généthon map, http://www.genethon.fr/genethon_en.html (for distal marker identification)
    1. Genome Database, http://gdbwww.gdb.org/ (for primer information)

References

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