Fig 1.

Targeted mutation of mouse NHERF-1 gene. (A) The NotI genomic fragments, A5 and B12, from the mouse NHERF-1 gene were used to design a targeting vector. PCR product encompassing the BstII/XbaI fragment from A5 and the NotI/KpnI fragment from B12 were ligated to Neo^r gene, which was combined with herpes simplex virus thymidylate kinase (HSV-TK; KpnI fragment) to construct a targeting vector. Primers based on the shared BstII/XbaI fragment (primer 1) and the Neo^r gene (primer 2) or the first exon of the mouse NHERF-1 gene (primer 3) were used to amplify the WT and mutant alleles by using the PCR. B, BstII; X, XbaI; K, KpnI; N, NotI; S, SacI (indicate the restriction sites). (B) Products generated by PCR amplification of genomic DNA from WT (+/+), heterozygous (+/−), and null (−/−) mice. (C) Southern blot of the BglII digest of mouse genomic DNA hybridized with the PCR product from the A5 region described in Materials and Methods.