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A 3.6 Kb BamHI fragment containing exon 2 was replaced with a neomycin selection cassette. Correct targeting was confirmed by Southern blotting. Analysis of mRNAs from homozygote B cells demonstrates exon 1 was spliced to exon 3. All potential ATG start codons upstream of exon 3 and located in the correct reading frame are followed by at least 2 stop codons prior to the start of exon 3. The first ATG present within the functional reading frame downstream of the start of exon 3 is located in exon 5, which could only generate a 157 amino acid long fragment of the 555 amino acid endogenous protein. For these reasons, a functional protein product was considered unlikely. (J:144320)