FIG. 5.

Targeted disruption of the N6amt1 gene in mice. (A) Targeting strategy to generate the mutant N6amt1 allele. Numbered gray boxes represent coding exons, and open boxes represent the 3′ untranscribed region (UTR). B, BamHI restriction site; Neo, neomycin resistance gene; HSV-tk, herpes simplex virus thymidine kinase gene. The locations of the Southern probe and the expected BamHI fragments are indicated. PCR primers used for genotyping are indicated with arrows. (B) Genotyping of ES clones by Southern blot analysis. Genomic DNA was digested with BamHI. The 9.5-kb and 5.7-kb bands were derived from the wild-type and targeted alleles, respectively. (C) Genotyping of mouse embryos by PCR. Heterozygous mice were intercrossed, and E3.5 blastocysts were collected. The 267-bp and 372-bp PCR products were derived from the wild-type and targeted alleles, respectively.