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A targeting vector was used to insert an inactivated muscle-specific 5' donor splice site and a strong polyA site. A neomycin resistance cassette replaced untranslated exons 23 and 24, and the muscle-specific exon 25. The resulting allele lacked expression of the protein in the heart and prevented its replacement by the b isoform. RNase protection assay and Western blot failed to detect transcript and protein, respectively, in mutants. (J:111247)