410 Nucleic Acids Research, Vol. 19, No. 2

An STS in the human T-cell receptor delta locus located at 14q11 P.Chuchana, F.Bernard, G.Lefranc and M.-P.Lefranc* Laboratoire d'lmmunogenetique Moleculaire, URA CNRS 1191, Universite Montpellier 11, CP 012, Place E. Bataillon, 34095 Montpellier Cedex 5, France EMBL accession no. X15207

Submitted October 10, 1990

The human T cell receptor delta locus (TCRD or TRD) located at 14q1 1 is embedded in the T-cell receptor alpha locus between the Va and the Ja segments at 85 kb upstream of the Cca gene (1). The human T-cell receptor delta locus comprises a unique constant region gene CA preceded by 3 J6 and 3 Db (for review, see ref. 2). The number of the variable VA genes seems limited. We isolated the first genomic sequence of the T- cell receptor V62 (TCRDV2) gene (3). We have defined this information gene as a sequence-tagged site (STS), designated TCRDV2[/14ql1], for inclusion in the human genome map. Using the polymerase chain reaction (PCR) described below, a fragment of the expected size (320 bp) was amplified from human genomic DNA. The fragment contained sequences from positions 192 to 511 of the human TCRDV2 gene (3). The amplified fragment was purified, uniformly radiolabelled, and used to probe a human genomic Southern. As anticipated, a single 4.0 kb HindH, 3.6 kb XbaI and 12.5 kb EcoRI fragment were detected (3), demonstrating both the unique character of the amplified sequences, and the direct utility of the PCR product as a probe for retrieving clones containing this STS from libraries. BamHI identifies three allelic restriction fragments of 14.0 kb (TCRDV2*A1), 10.8 kb (*A2) and 14.5 kb (*A3) (4).

(Perkin Elmer Cetus) in 100 tl of 1 x PCR buffer (50 mM KCl, 10 mM Tris-HCl, pH8.3 (at room temperature), 1.5 mM MgCl2, 0.1% (w/v) gelatin).

PCR profile: 94°C for 5 minutes 60°C for 10 minutes 720C for 1 minute 940C for 1 minute for 30 cycles 600C for 1 minute Anticipated sequence of the PCR product: CTCAGGAGTCATGTCAGCCATTGAGTTGGTGCCTGAACACCAAACAGTGCCTGTGTCAAT AGGGGTCCCTGCCACCCTCAGGTGCTCCATGAAAGGAGAAGCGATCGGTAACTACTATAT CAACTGGTACAGGAAGACCCAAGGTAACACAATCACTTTCATATACCGAGAAAAGGACAT CTATGGCCCTGGTTTCAAAGACAATTTCCAAGGTGACATTGATATTGCAAAGAACCTGGC TGTACTTAAGATACTTGCACCATCAGAGAGAGATGAAGGGTCTTACTACTGTGCCTGTGA CACCCACCCTGCTGCAGCTC

PCR primers: Forward (192-210 in ref. 3) CTCAGGAGTCATGTCAGCC Reverse (511-494 in ref. 3) GAGCTGCAGGGTGGG PCR components: 2 yg of human genomic DNA, 40 pmoles of each nucleotide, 200 jtM dNTPs, and 2.5 U Taq polymerase

REFERENCES 1. 2. 3. 4.

Boehm,T. et al. EMBO J. 7, 385-394, (1988). Lefranc,M.-P. Eur. Cytokine Net. 1, 121-130 (1990). Dariavach,P. and Lefranc,M.P. Nucl. Acids Res. 17, 4880 (1989). Chuchana,P. et al. Nucl. Acids Res. 17, 3622 (1989).

An STS in the human T-cell receptor delta locus located at 14q11.

410 Nucleic Acids Research, Vol. 19, No. 2 An STS in the human T-cell receptor delta locus located at 14q11 P.Chuchana, F.Bernard, G.Lefranc and M.-P...
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