Summary
The construction of an extensive human genetic linkage map will require the generation of large numbers of DNA probes specific for single chromosomes. Recombinant DNA libraries representing chromosomes 22 and 21 have been constructed with a view to studying the specific rearrangements of chromosome 22 observed in chronic myeloid leukaemia and Burkitt’s lymphoma and also the association of chromosome 21 with Down’s syndrome. This was accomplished by sorting about 2 × 106 copies of chromosome 22 and 21 by flow cytometry and insertion of the DNA into the vector λgt WESλB. Twenty clones selected at random from the chromosome 22 library hybridised to EcoRI-digested human DNA, and five of these hybridised to single bands identical in size to the phage inserts. Altogether six single-copy sequences and a clone coding for an 8S RNA isolated by screening the chromosome 22 library for expressed sequences were characterised in detail. Hybridisation of all seven clones to a panel of sorted chromosomes and hybrid cell lines confirmed the assignment of the sequences to chromosome 22. The sequences were localised to regions of chromosome 22 by hybridisation 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 qter, while two clones and an 18S rRNA gene isolated from the chromosome 22 library reside between pter and q112. In situ hybridisation has further mapped one of these clones to the region q13-qter. An identical approach was used for chromosome 21 and three out of five single copy clones have been identified as specific to this chromosome by hybridisation to DNA from a cell hybrid containing only human chromosome 21. In summary, it has been shown that this approach has yielded DNA libraries of high purity based on chromosomes 21 and 22. This method has the advantage of being direct and applicable to nearly all human chromosomes and will be important in the molecular analyses of human genetic disease.
This work was supported by the Medical Research Council, Cancer Research Campaign and the Leukemia Research Fund
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Similar content being viewed by others
References
Ruddle FH (1981) Nature 294: 115–120
Padgett TF, Stubblefield E, Varmus HE (1977) Cell 10: 649–657
Pinaev G, Bardyopadhyay D, Glekov O, Shanbag V, Johanson G, Albertson PA (1979) Exp Cell Res 124: 191–203
Collard JG, Schijven J, Tulp A, Meulenbrock M (1982) Exp Cell Res 137:463–469
McKusick VA, Ruddle FH (1977) Science 196: 390–405
Gusella J, Keys C, Varsanyi-Breiner A, Kao F, Jones C, Puck TT, Housman D (1980) Proc Natl Acad Sci USA 77:2829–2333
Krumlauf R, Jeanpierre M, Young BD (1982) Proc Natl Acad Sci USA 79:2971–2975
Sillar RS, Young BD (1981) J Histochem Cytochem 29:74–78
Carrano AV, Gray JW, Langlois RG, Burkhart-Schultz K, Van Dilla MA (1979) Proc Natl Acad Sci USA 76: 1382–1384
Young BD, Ferguson-Smith MA, Sillar RS, Boyd E (1981) Proc Natl Acad Sci USA 78:7727–7731
Maniatis T, Hardison RC, Lowry E, Lauer J, O’Connell C, Quon D, Sim GK, Efstradiatis A (1978) Cell 15:687–701
Rigby RWJ, Diekmann M, Rhodes C, Berg P (1977) J Mol Biol 113:237–254
Schmidt CW, Deininger PL (1975) Cell 6:345–358
Lebo RV, Carrano AV, Burkhart-Schultz K, Dozy AM, Yu LC, Kan YW (1979) Proc Natl Acad Sci USA 76:5804–5808
Malcolm S, Williamson R, Boyd E, Ferguson-Smith MA (1977) Cytogenet Cell Genet 19:256–261
Wolf SF, Mareni C, Migeon B (1980) Cell 21:95–102
Erikson J, Martinis J, Croce CM (1981) Nature 294: 173–175
Klein G (1981) Nature 294:313–318
Davies KE, Young BD, Elles RG, Hill ME, Williamson R (1981) Nature 293:374–376
Kunkel LM, Tantravahi U, Eisenhard M, Latt SA (1982) Nucl Acids Res 10: 1557–1578
Disteche CM, Kunkel LM, Lojewski A, Orkin SH, Eisenhard M, Sahar E, Travis B, Latt SA (1982) Cytometry 2:282–286
Gray JW, Langlois RG, Carrano A V, Burkhart-Schultz K, Van Dillen MA (1979) Chromosoma 73:9–27
Rowley JD (1973) Nature 243:290–293
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1983 Springer-Verlag Berlin Heidelberg
About this paper
Cite this paper
Young, B.D., Jeanpierre, M., Goyns, M.H., Stewart, G.D., Elliot, T., Krumlauf, R. (1983). Construction and Characterization of Chromosomal DNA Libraries. In: Neth, R., Gallo, R.C., Greaves, M.F., Moore, M.A.S., Winkler, K. (eds) Modern Trends in Human Leukemia V. Haematology and Blood Transfusion / Hämatologie und Bluttransfusion, vol 28. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-68761-7_60
Download citation
DOI: https://doi.org/10.1007/978-3-642-68761-7_60
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-540-11858-9
Online ISBN: 978-3-642-68761-7
eBook Packages: Springer Book Archive