Abstract
Yeast artificial chromosomes (YACs) were used to construct a physical map of the germline human T-cell β chain gene complex (TCRB). Variable region genes (BV) for the 25 known subfamilies were used as probes to screen the ICRF AM4x YAC library. Of the five positive YACs identified, one YAC designated B3, 820 kilobase pairs (kbp) in size, scored positive for all 25 TCRBV subfamilies plus the constant region genes (BC) when analyzed by pulse field gel electrophoresis. Restriction enzyme mapping of B3 located TCRBV and TCRBC gene regions to 4 Sfi I fragments of 280 110, 90, and 125 kbp and was in accordance with published data. In addition comparison of hybridization results of Sfi I-restricted B3 and genomic DNA from the parental cell line GM1416B revealed identical banding patterns. The data thus showed YAC B3 encoded a complete and unrearranged TCRB gene locus of some 600–620 kbp. The map was further resolved by locating restriction sites for Sal I and Bss HII on B3, giving more precise localization of the individual TCRBV gene families. Flourescent in situ hybridization of B3 to spreads of human metaphase chromosomes localized B3 to 7q35. However, two additional signals were obtained; one attributable to the TCRBV orphon cluster on 9p21, the second to the long arm of chromosome 2. Polymerase chain reaction amplification of a chromosome 2 somatic cell hybrid, using primers for all 25 TCRBV gene families, revealed that the signal was not attributable to a second orphon cluster. It is suggested that B3 is a chimeric YAC with an intact TCRB locus flanked by chromosome 2 sequences.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Anand, R., Ogilvie, D. J., Butler, R., Riley, J. H., Finniear, R. S., Powell, S. J., Smith, J. C., and Markham, A. F. A yeast artificial chromosome contig encompassing the cystic fibrosis locus. Genomics 9: 124–130, 1991
Blin, N. and Stafford, D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res 3: 2303, 1976
Bonneau, D., Huret, J. L., Godeau, G., Couet, D., Putterman, M., Tanzer, J., Babin, P., and Larregue, M. Recurrent ctb(7) (q31.3) and possible laminin involvement in a neonatal cutis laxa with a Marfan phenotype. Hum Genet 87: 317–319, 1991
Buckle, V. J. and Rack, K. Fluorescent in situ hybridization. In K. E. Davies (ed.): Human Genetic Disease Analysis: A Practical Approach, vol 2, pp. 59–82. IRL Press, Oxford, 1993
Burke, D. T., Carle, G. F., and Olson, M. V. Cloning of large DNA segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science 236: 806–812, 1987
Charmley, P., Wei, S., and Concannon, P. Polymorphisms in the Tcrb-V2 gene segments localize the Tcrb orphon genes to human chromosome 9p21. Immunogenetics 38: 283–286, 1993
Childs, G., Maicson, R., Cohn, R. H., and Kedes, L. Orphons: dispersed genetic elements derived from tandem repetitive genes of eukaryotes. Cell 23: 651–653, 1988
Choi, K., Kotzin, B., Heron, L., Callahan, J., Marrack, P., and Keppler, J. Interaction of Staphylococcus aureus toxins superantigens with human T-cells. Proc Natl Acad Sci USA 86: 8941–8949, 1989
Concannon, P., Pickering, L. A., Kung, P., and Hood, L. Diversity and structure of human T-cell receptor β-chain variable region genes. Proc Natl Acad Sci USA 83: 6598–6602, 1986
Davis, M. M. and Bjorkman, P. J. T-cell receptor genes and T-cell recognition. Nature 334: 395–398, 1988
Feinberg, A. P. and Vogelstein, B. A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6, 1983
Garza, D., Ajioka, J. W., Burke, D. T., and Hartl, D. L. Mapping the Drosophila genome with yeast artificial chromosomes. Science 246: 641–646, 1989
Green, E. D. and Olson, M. V. Systematic screening of yeast artificial chromosome libraries by use of the polymerase chain reaction. Proc Natl Acad Sci USA 87: 1213–1217, 1990
Gurrieri, F., Trask, B. J., van den Engh, G., Krauss, C. M., Schinzel, A., Pettenati, M. J., Schindler, D., Dietz-band, J., Vergnaud, G., Scherer, S. W., and Tsui, L.-C. Physical mapping of the holoprosencephaly critical region on chromosome 7q36. Nature Genetics 3: 247–251, 1993
Hall, M. A., Jaeger, E. E. M., Boutrop, R. E., and Lanchbury, J. S. Characterization of a novel human T-cell receptor β chain variable region family by transspecies DNA hybridization. Eur J Immunol 24: 641–645, 1994
Huber, C., Thiebe, R., Hamiester, H., Bromola, H., Lotscher, E., and Zachau, H. G. A human immunoglobulin kappa orphon without sequence defects may be the product of a pericentric inversion. Nucleic Acids Res 18: 3475–3478, 1990
Isobe, M., Erickson, J., Emanuel, B. S., Nowell, P. C., and Croce, C. M. Location of gene for β subunit of human T-cell receptor at band 7q35, a region prone to rearrangements in T cells. Science 228: 580–582, 1985
Kere, J., Davies, K. A., Roninson, I. B., Watkins, P. C., Winqvist, R., and de la Çhapelle, A. Chromosome 7 long arm deletion in myeloid disorders. A narrow breakpoint region in 7q22 defined by molecular mapping. Blood 73: 230–234, 1989
Lai, E., Concannon, P., and Hood, L. Conserved organisation of the human and murine T-cell receptor beta gene families. Nature 331: 543–546, 1988
Laring, Z., Monaco, A., and Lehrach, H. Yeast artificial chromosome libraries containing large inserts from mouse and human DNA. Proc Natl Acad Sci USA 8: 4123–4127, 1991
Marrack, P. and Kappler, K. The T-cell receptor. Science 238: 1073, 1987
Robinson, M. A., Mitchell, M. P., Wei, S., Day, C., Zhao, T., and Concannon, P. Organisation of human T-cell receptor β-chain genes: clusters of Vβ genes are present on chromosomes 7 and 9. Proc Natl Acad Sci USA 90: 2433–2437, 1993
Seboun, E., Robinson, M. A., Kindt, T. J., and Hauser, S. L. Insertion/deletion-related polymorphisms in the human T cell receptor β gene complex. J Exp Med 170: 1263–1270, 1989
Seboun, E., Houghton, C. J., Hatem, C. J., Lincoln, R., and Hauser, S. L. Unusual organisation of the human T-cell receptor β-chain gene complex is linked to recombination hotspots. Proc Natl Acad Sci USA 90: 5026–5029, 1993
Sharland, M., Patton, M., and Hill, L. Ectrodactyly of hands and feet in a child with a complex translocation including 7q21.2. Am J Med Genet 38: 413–414, 1991
Slightom, J. L., Siemieniak, D. R., Sieu, L. C., Koop, B. F., and Hood, L. Nucleotide sequence analysis of 77.7 kb of the human Vb T-cell receptor gene locus: direct primer-walking using cosmid template DNAs. Genomics 20: 149–168, 1994
Toyonaga, B., Yoshikai, Y., Vadsaz, V., Chin, B., and Mak, T. W. Organisation and sequences of the diversity, joining and constant region genes of the human T-cell receptor β-chain. Proc Natl Acad Sci USA 82: 8624, 1985
Toyonaga, B. and Mak, T. W. Genes of the T-cell antigen receptor in normal and malignant T cells. Annu Rev Immunol 5: 585–620, 1987
Van Ommen, G. J. B. and Verkerk, J. M. Restriction analysis of chromosomal DNA in a size range up to two million base pairs by pulse field gradient electrophoresis. In K. E. Davis (ed.): Human Genetic Disease: A Practical Approach, pp. 113–133, IRL Press, Oxford, 1986
Wei, S., Charmley, P., Robinson, M. A., and Concannon, P. The extent of the human germline T-cell receptor V beta gene segment repertoire. Immunogenetics 40: 27–36, 1994
Wilson, R. K., Lai, E., Concannon, P., Barth, R. K., and Hood, L. E. Structure, organisation and polymorphism of murine and human T-cell receptor alpha and beta chain gene families. Immunol Rev 101: 149–172, 1988
WHO-IUIS Nomenclature Sub-Committee on TCR Designation: Nomenclature for T-cell receptor (TCR) gene segments of the immune system. Eur J Immunol 23: 2729–2731, 1993
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hashim, Y., Ragoussis, I., Kearney, L. et al. Physical mapping of the human T-cell receptor beta gene complex, using yeast artificial chromosomes. Immunogenetics 41, 337–342 (1995). https://doi.org/10.1007/BF00163990
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00163990