Physical mapping of the human t-cell receptor beta gene complex, using yeast artificial chromosomes

Journal Article · Sat Apr 01 04:00:00 EST 1995 · Immunogenetics

DOI:https://doi.org/10.1007/BF00163990· OSTI ID:91083

Hashim, Y; So, A K ^[1]; Kearney, L ^[2]

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1. Hammersmith Hospital, London (United Kingdom)
2. Institute of Molecular Medicine, Oxford (United Kingdom); and others

Yeast artificial chromosomes (YACs) were used to construct a physical map of the germline human T-cell {beta} 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. Fluorescent 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. 32 refs., 5 figs., 1 tab.

• Sponsoring Organization: USDOE
• OSTI ID: 91083
• Journal Information: Immunogenetics, Journal Name: Immunogenetics Journal Issue: 6 Vol. 41; ISSN 0093-7711; ISSN IMNGBK
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
ANIMAL CELLS
CHIMERAS
DNA HYBRIDIZATION
FLUORESCENCE
GENES
GENETIC MAPPING
HUMAN CHROMOSOME 2
HUMAN CHROMOSOME 7
HUMAN CHROMOSOME 9
HUMAN CHROMOSOMES
IMMUNITY
RECEPTORS
SIZE
YEASTS