Cloning and sequencing of the mouse Capn2 locus
The mouse Capn2 gene encodes the 700 amino acids of the m-calpain large subunit (m-80 k) and consists of 21 exons extending over 50-kb on chromosome 1. A cDNA clone encoding a portion of the mouse m-80 k subunit was purchased (dbEST Id, 807416, Image:606689, Image Consortium, LLNL). It was found to contain 2.8-kb of sequence from position 247 of the coding sequence to the stop codon at position 2,101, including the 3'-UTR to the polyA signal. A fragment of this cDNA was subcloned and used to screen a 129Sv mouse genomic library in λ-Dash II. Overlapping genomic clones were obtained covering 15,354 bp, extending from a BamHI site in intron 3 to a BamHI site in intron 15. These clones were sequenced completely on both strands and the data were submitted to GenBank (accession no. AF497625). The submitted sequences agreed precisely with the public databases, and also filled in several small gaps corresponding to short repetitive sequences which could only be firmly established by repeated sequencing in non-standard conditions.