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PMID: 9892678 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Human artificial chromosomes generated by modification of a yeast artificial chromosome containing both human alpha satellite and single-copy DNA sequences.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 2 ·1999-01-19 ·Pages 592-7

Henning KA, Novotny EA, Compton ST, Guan XY, Liu PP, Ashlock MA

Abstract

A human artificial chromosome (HAC) vector was constructed from a 1-Mb yeast artificial chromosome (YAC) that was selected based on its size from among several YACs identified by screening a randomly chosen subset of the Centre d'Etude du Polymorphisme Humain (CEPH) (Paris) YAC library with a degenerate alpha satellite probe. This YAC, which also included non-alpha satellite DNA, was modified to contain human telomeric DNA and a putative origin of replication from the human beta-globin locus. The resultant HAC vector was introduced into human cells by lipid-mediated DNA transfection, and HACs were identified that bound the active kinetochore protein CENP-E and were mitotically stable in the absence of selection for at least 100 generations. Microdissected HACs used as fluorescence in situ hybridization probes localized to the HAC itself and not to the arms of any endogenous human chromosomes, suggesting that the HAC was not formed by telomere fragmentation. Our ability to manipulate the HAC vector by recombinant genetic methods should allow us to further define the elements necessary for mammalian chromosome function.

MeSH Terms
Cell Line Chromosomes/genetics Chromosomes, Artificial, Yeast/genetics Clone Cells/metabolism DNA Fragmentation/genetics DNA Probes/genetics DNA, Satellite/genetics Gene Dosage Genetic Vectors/genetics Globins/genetics Humans In Situ Hybridization, Fluorescence Mitosis/genetics Replication Origin/genetics Telomere/genetics Transfection/genetics
Chemicals
DNA Probes DNA, Satellite Globins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Henning K A
Genetics and Molecular Biology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Novotny E A
Compton S T
Guan X Y
Liu P P
Ashlock M A
References (48)
48 references, click to expand
  1. The CEPH YAC library.
    Behring Inst Mitt. 1992 Apr;(91):13-20 PMID: 1524562
  2. Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer.
    Genomics. 1992 Jul;13(3):718-25 PMID: 1639399
  3. Detection and characterization of chimeric yeast artificial-chromosome clones.
    Genomics. 1991 Nov;11(3):658-69 PMID: 1723055
  4. Functional reintroduction of human telomeres into mammalian cells.
    Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7006-10 PMID: 1871116
  5. Labeling of the centromeric region on human chromosome 8 by in situ hybridization.
    Hum Genet. 1991 Aug;87(4):489-94 PMID: 1879835
  6. A novel, rapid method for the isolation of terminal sequences from yeast artificial chromosome (YAC) clones.
    Nucleic Acids Res. 1990 May 25;18(10):2887-90 PMID: 2161516
  7. Structural instability of human tandemly repeated DNA sequences cloned in yeast artificial chromosome vectors.
    Nucleic Acids Res. 1990 Mar 25;18(6):1421-8 PMID: 2183192
  8. Structure and polymorphism of human telomere-associated DNA.
    Cell. 1990 Oct 5;63(1):119-32 PMID: 2208276
  9. Rapid isolation of human chromosome-specific DNA probes from a somatic cell hybrid.
    Genomics. 1990 Jun;7(2):257-63 PMID: 2347589
  10. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  11. Chromosome-specific organization of human alpha satellite DNA.
    Am J Hum Genet. 1985 May;37(3):524-32 PMID: 2988334
  12. Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors.
    Science. 1987 May 15;236(4803):806-12 PMID: 3033825
  13. A subfamily of alphoid repetitive DNA shared by the NOR-bearing human chromosomes 14 and 22.
    Genomics. 1988 Aug;3(2):100-9 PMID: 3224978
  14. Chromosome segregation in mitosis and meiosis.
    Annu Rev Cell Biol. 1985;1:289-315 PMID: 3916318
  15. Construction of artificial chromosomes in yeast.
    Nature. 1983 Sep 15-21;305(5931):189-93 PMID: 6350893
  16. Chromosomal localization of complex and simple repeated human DNAs.
    Chromosoma. 1978 Mar 22;66(1):23-32 PMID: 639625
  17. The molecular genetics of human hemoglobin.
    Prog Nucleic Acid Res Mol Biol. 1984;31:315-462 PMID: 6397774
  18. Centromere activation.
    Trends Genet. 1995 Sep;11(9):337-9 PMID: 7482782
  19. Molecular cloning of a novel protein regulated by opioid treatment of NG108-15 cells.
    Brain Res Mol Brain Res. 1995 Aug;32(1):171-5 PMID: 7494458
  20. A human telomeric protein.
    Science. 1995 Dec 8;270(5242):1663-7 PMID: 7502076
  21. Further evidence that CENP-C is a necessary component of active centromeres: studies of a dic(X; 15) with simultaneous immunofluorescence and FISH.
    Hum Mol Genet. 1995 Feb;4(2):289-94 PMID: 7757082
  22. Dissecting the centromere of the human Y chromosome with cloned telomeric DNA.
    Hum Mol Genet. 1994 Aug;3(8):1227-37 PMID: 7987296
  23. Addition of functional human telomeres to YACs.
    Hum Mol Genet. 1994 Aug;3(8):1383-6 PMID: 7987319
  24. Characterization of four human YAC libraries for clone size, chimerism and X chromosome sequence representation.
    Nucleic Acids Res. 1994 Aug 25;22(16):3406-11 PMID: 8078777
  25. Stringent sequence requirements for the formation of human telomeres.
    Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8861-5 PMID: 8090736
  26. Initiation of chromosomal DNA replication in eukaryotes. Lessons from lambda.
    J Biol Chem. 1994 Mar 11;269(10):7047-50 PMID: 8125908
  27. Replication structure of the human beta-globin gene domain.
    Nature. 1993 Dec 9;366(6455):588-90 PMID: 8255298
  28. NIGMS human/rodent somatic cell hybrid mapping panels 1 and 2.
    Genomics. 1993 May;16(2):311-4 PMID: 8314568
  29. Characterization of NIGMS human/rodent somatic cell hybrid mapping panel 2 by PCR.
    Genomics. 1993 May;16(2):315-9 PMID: 8314569
  30. Microinjection of intact 200- to 500-kb fragments of YAC DNA into mammalian cells.
    Genomics. 1993 Mar;15(3):659-67 PMID: 8385648
  31. Generation of band-specific painting probes from a single microdissected chromosome.
    Hum Mol Genet. 1993 Aug;2(8):1117-21 PMID: 8401492
  32. Telomere directed fragmentation of mammalian chromosomes.
    Nucleic Acids Res. 1993 Jan 11;21(1):27-36 PMID: 8441617
  33. DUB-1, a deubiquitinating enzyme with growth-suppressing activity.
    Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3275-9 PMID: 8622927
  34. Identification of centromeric antigens in dicentric Robertsonian translocations: CENP-C and CENP-E are necessary components of functional centromeres.
    Hum Mol Genet. 1995 Dec;4(12):2189-97 PMID: 8634687
  35. The kinesin-like protein CENP-E is kinetochore-associated throughout poleward chromosome segregation during anaphase-A.
    J Cell Sci. 1996 May;109 ( Pt 5):961-9 PMID: 8743943
  36. A method for linking yeast artificial chromosomes.
    Nucleic Acids Res. 1996 Nov 1;24(21):4192-6 PMID: 8932371
  37. Functional human CFTR produced by stable Chinese hamster ovary cell lines derived using yeast artificial chromosomes.
    Hum Mol Genet. 1997 Jan;6(1):59-68 PMID: 9002671
  38. Formation of de novo centromeres and construction of first-generation human artificial microchromosomes.
    Nat Genet. 1997 Apr;15(4):345-55 PMID: 9090378
  39. A functional neo-centromere formed through activation of a latent human centromere and consisting of non-alpha-satellite DNA.
    Nat Genet. 1997 Jun;16(2):144-53 PMID: 9171825
  40. Human centromeric DNAs.
    Hum Genet. 1997 Sep;100(3-4):291-304 PMID: 9272147
  41. Contiguous arrays of satellites 1, 3, and beta form a 1.5-Mb domain on chromosome 22p.
    Genomics. 1997 Aug 15;44(1):35-44 PMID: 9286698
  42. Telomeric localization of TRF2, a novel human telobox protein.
    Nat Genet. 1997 Oct;17(2):236-9 PMID: 9326951
  43. Centromere DNA dynamics: latent centromeres and neocentromere formation.
    Am J Hum Genet. 1997 Dec;61(6):1225-33 PMID: 9399915
  44. TRF2 protects human telomeres from end-to-end fusions.
    Cell. 1998 Feb 6;92(3):401-13 PMID: 9476899
  45. Humanizing the yeast telomerase template.
    Proc Natl Acad Sci U S A. 1998 May 12;95(10):5667-71 PMID: 9576941
  46. Centromeres take flight: alpha satellite and the quest for the human centromere.
    Cell. 1998 May 1;93(3):317-20 PMID: 9590164
  47. Construction of YAC-based mammalian artificial chromosomes.
    Nat Biotechnol. 1998 May;16(5):431-9 PMID: 9592390
  48. Genetic dissection of a mammalian replicator in the human beta-globin locus.
    Science. 1998 Aug 14;281(5379):1005-9 PMID: 9703500
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.2.592
Published
1999-01-19
Pages
592-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC15181
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.96.2.592 PMC Full Text View journal details
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