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. 1999 Oct 18;190(8):1093–1102. doi: 10.1084/jem.190.8.1093

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© 1999 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Thymocytes from TCR-α/β–transgenic SLP-76^−/− mice fail to suppress V to DJ rearrangements in the endogenous TCR-β locus. 20 ng of genomic DNA isolated from DN thymocytes of SLP-76^+/−, SLP-76^+/−-tg, SLP-76^−/−, and SLP-76^−/−-tg was amplified with primers that specifically detect rearrangements among Vβ5, Vβ8, Vβ10, and DJβ2. Equivalent loading was confirmed by amplification of a fragment of the Cμ gene (IgM). The PCR products were separated on a 1.5% agarose gel, transferred to a nylon membrane, and hybridized with radiolabeled oligonucleotide probes specific for Jβ2.7 and Cμ. For each Vβ, there are six bands that correspond to rearrangements to the DJβ2 segments DJβ2.1 through DJβ2.7 (Jβ2.6 is a pseudogene and is not rearranged). Similar results were obtained in two other experiments.