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PMID: 6953442 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Construction and characterization of genomic libraries from specific human chromosomes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 9 ·1982-05-00 ·Pages 2971-5

Krumlauf R, Jeanpierre M, Young BD

Abstract

Highly purified fractions of human chromosomes 21 and 22 were isolated from a suspension of metaphase chromosomes stained with ethidium bromide by using a fluorescence-activated cell sorter (FACS II). Two recombinant DNA libraries, representing chromosomes 21 and 22, were constructed by complete digestion of DNA from these fractions with EcoRI and insertion into the vector lambda gtWES lambda B. Twenty clones selected at random from the chromosome 22 library hybridized to EcoRI-digested human DNA, and five of these clones hybridized to single bands identical in size to the phage inserts. These five single-copy sequences and a clone coding for an 8S RNA isolated by screening the chromosome 22 library for expressed sequences were characterized in detail. Hybridization of all six clones to a panel of sorted chromosomes and hybrid cell lines confirmed the assignment of the sequences to chromosome 22. The sequences were localized to regions of chromosome 22 by hybridization to translocated chromosomes sorted from a cell line having a balanced translocation t(17;22)(p13;q11) and to hybrid cell lines containing the various portions of another translocation t(X;22)(q13;q112). Five clones reside on the long arm of chromosome 22 between q112 and pter, while one clone and an 18S rRNA gene isolated from the chromosome 22 library reside pter and g112. The construction of chromosome-specific libraries by this method has the advantage of being direct and applicable to nearly all human chromosomes and will be important in molecular analysis of human genetic diseases.

MeSH Terms
Cell Line Chromosome Mapping/methods Chromosomes, Human Chromosomes, Human, 21-22 and Y Cloning, Molecular/methods DNA, Recombinant Flow Cytometry/methods Humans
Chemicals
DNA, Recombinant
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Krumlauf R
Jeanpierre M
Young B D
References (28)
28 references, click to expand
  1. Isolation of high-molecular-weight DNA from mammalian cells.
    Eur J Biochem. 1973 Jul 2;36(1):32-8 PMID: 4200179
  2. Letter: A new consistent chromosomal abnormality in chronic myelogenous leukaemia identified by quinacrine fluorescence and Giemsa staining.
    Nature. 1973 Jun 1;243(5405):290-3 PMID: 4126434
  3. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  4. EK2 derivatives of bacteriophage lambda useful in the cloning of DNA from higher organisms: the lambdagtWES system.
    Science. 1977 Apr 8;196(4286):175-7 PMID: 322278
  5. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.
    J Mol Biol. 1977 Jun 15;113(1):237-51 PMID: 881736
  6. Application of endonuclease mapping to the analysis and prenatal diagnosis of thalassemias caused by globin-gene deletion.
    N Engl J Med. 1978 Jul 27;299(4):166-72 PMID: 661890
  7. Sequence organization of the human genome.
    Cell. 1975 Nov;6(3):345-58 PMID: 1052772
  8. The isolation of structural genes from libraries of eucaryotic DNA.
    Cell. 1978 Oct;15(2):687-701 PMID: 719759
  9. Polymorphism of DNA sequence adjacent to human beta-globin structural gene: relationship to sickle mutation.
    Proc Natl Acad Sci U S A. 1978 Nov;75(11):5631-5 PMID: 281713
  10. Measurement and purification of human chromosomes by flow cytometry and sorting.
    Proc Natl Acad Sci U S A. 1979 Mar;76(3):1382-4 PMID: 286320
  11. Partial deletion of beta-globin gene DNA in certain patients with beta 0-thalassemia.
    Proc Natl Acad Sci U S A. 1979 May;76(5):2400-4 PMID: 287080
  12. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate.
    Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683-7 PMID: 291033
  13. DNA sequence variants in the G gamma-, A gamma-, delta- and beta-globin genes of man.
    Cell. 1979 Sep;18(1):1-10 PMID: 509514
  14. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.
    Biochemistry. 1979 Nov 27;18(24):5294-9 PMID: 518835
  15. Assignment of human beta-, gamma-, and delta-globin genes to the short arm of chromosome 11 by chromosome sorting and DNA restriction enzyme analysis.
    Proc Natl Acad Sci U S A. 1979 Nov;76(11):5804-8 PMID: 293684
  16. A new translocation in Burkitt's tumor cells.
    Hum Genet. 1979;53(1):111-2 PMID: 535896
  17. Model for antenatal diagnosis of beta-thalassaemia and other monogenic disorders by molecular analysis of linked DNA polymorphisms.
    Nature. 1980 May 15;285(5761):144-7 PMID: 6246441
  18. Construction of a genetic linkage map in man using restriction fragment length polymorphisms.
    Am J Hum Genet. 1980 May;32(3):314-31 PMID: 6247908
  19. Isolation and localization of DNA segments from specific human chromosomes.
    Proc Natl Acad Sci U S A. 1980 May;77(5):2829-33 PMID: 6930670
  20. Isolation and characterization of cloned DNA sequences that hybridize to the human X chromosome.
    Cell. 1980 Aug;21(1):95-102 PMID: 7190879
  21. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201-5 PMID: 6159641
  22. A highly polymorphic locus in human DNA.
    Proc Natl Acad Sci U S A. 1980 Nov;77(11):6754-8 PMID: 6935681
  23. A new method for the preparation of metaphase chromosomes for flow analysis.
    J Histochem Cytochem. 1981 Jan;29(1):74-8 PMID: 6162882
  24. Cloning of a representative genomic library of the human X chromosome after sorting by flow cytometry.
    Nature. 1981 Oct 1;293(5831):374-6 PMID: 6456416
  25. Regional assignment of arylsulfatase A, mitochondrial aconitase and NADH-cytochrome b5 reductase by somatic cell hybridization.
    Hum Genet. 1981;58(2):140-3 PMID: 6116664
  26. Assignment of the genes for human lambda immunoglobulin chains to chromosome 22.
    Nature. 1981 Nov 12;294(5837):173-5 PMID: 6795508
  27. The role of gene dosage and genetic transpositions in carcinogenesis.
    Nature. 1981 Nov 26;294(5839):313-8 PMID: 7312030
  28. High-resolution analysis of human peripheral lymphocyte chromosomes by flow cytometry.
    Proc Natl Acad Sci U S A. 1981 Dec;78(12):7727-31 PMID: 6950411
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.79.9.2971
Published
1982-05-00
Pages
2971-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC346330
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.79.9.2971 PMC Full Text View journal details
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