Tg(TcraCWM5,TcrbCWM5)1807Wuth

The CW/M 5 CD4+ T cell line is from C57BL/6 mice injected with cell wall/membrane (CW/M) of Blastomyces dermatitidis yeast cells. The identity of the T cell receptor (TCR) Valpha and Vbeta chains on the surface of the "CW/M 5" T cells was found to be Valpha2 Jalpha49 (amino acid motif of the CDR3 region is AGGYQNFYFGKGSLTVIP) and Vbeta1/D/Jbeta1.1 (amino acid motif of the CDR3 region is SQEGTGG). The TCR-alpha gene segment, containing the rearranged VJ region from 7 bp upstream of the start codon to 71 bp beyond the Jalpha49 coding region into the JC intron, was amplified from genomic DNA of clone #5. This fragment was cloned into the pTalpha cassette. In a similar manner, the TCR-beta gene segment containing the rearranged VDJ region from 16 bp upstream of the start codon to 17 bp beyond the Jbeta1.1 coding regions in the JC intron, was cloned into the pTbeta cassette. The resulting constructs were co-microinjected into C57BL/6 oocytes. Linkage PCR determined that the transgenes in line 1807 had co-integrated in a TCRbeta-TCRalpha head-to-tail and TCRalpha-TCRbeta tail-to-tail fashion. (J:182207)