Lcp2^{tm5(Lat/Lcp2)Gak}

An allele expressing a chimeric molecule containing the membrane and lipid raft-targeting sequences of LAT fused to the full-length Lcp2 (SLP-76) was created in the following manner. Base pairs 1-105 of murine Lat cDNA were fused to exon 1 and intron 1 of Lcp2. The ATG start site of the Lcp2 gene was mutated and the entire construct was placed next to a FRT-flanked neomycin cassette. After homologous recombination, mice carrying the correctly targeted allele were crossed with FLPe transgenic mice to remove the neomycin cassette. Immunoblot analysis of heterozygote thymocytes detected the 80 kDA chimeric protein and the 76 kDa wild-type protein. (J:147721)