Gene targeting strategy for mouse CD59a.

(A) Partial restriction map of the CD59a gene fragment and construction of the targeting vector. N, NotI; X, XbaI; E,EcoRI; B, BamHI; Xh, XhoI, H,HindIII. After correct targeting, a 7.6-kb fragment containing CD59a exon 2 should be deleted. Exons are represented by thick vertical bars. (B) Representative Southern blot analysis of wild-type (+/+), heterozygous (+/−) and knockout (−/−) mouse genomic DNA after HindIII digestion. The probe used was a 282-bp cDNA fragment specific to CD59a exon 4. The targeted allele was identified by the 5.9-kb band while the wild-type allele was indicated by the 12.6-kb band.