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. 1999 Aug 3;96(16):9142–9147. doi: 10.1073/pnas.96.16.9142

Figure 1.

Figure 1

Ugcg gene targeting. (A) The biosynthetic pathways for sphingolipids. Glucosylceramide is produced by the modification of ceramide by glucosylceramide synthase. Glucosylceramide is the core structure for the synthesis of higher-order GSLs. Ceramide also is used for synthesis of sphingomyelin and galactosylceramide. (B) The structure of the pUgcgNeo targeting vector is shown at the top. The wild-type Ugcg locus is in the middle. The structure of the targeted locus (UgcgΔEX7Neo) is on the bottom. The UgcgΔEX7Neo locus was detected by Southern blot analysis of genomic DNA after digestion with HpaI and ClaI and hybridization with the probe indicated. The wild-type allele yielded a 12.6-kb fragment, and the UgcgΔEX7Neo allele yielded a 7.4-kb fragment. The Southern blots were from tail DNA of wild-type (+/+), and UgcgΔEX7Neo/+ (+/−) mice derived from targeted ES cells. (C) The structure of the pUgcgHygro targeting vector is shown at the top. The wild-type Ugcg locus is in the middle. The structure of the targeted allele, UgcgEX7Hygro, is on the bottom. The UgcgEX7Hygro locus was detected by Southern blot analysis of genomic DNA after digestion with HpaI and ClaI and hybridization with the probe indicated. The wild-type allele yielded a 12.6-kb band, and the UgcgΔEX7Neo and UgcgΔEX7Hygro alleles each yielded a 7.4-kb fragment. The Southern blots were of DNA of UgcgΔEX7Neo (+/−) ES cells and doubly targeted ES cells UgcgΔEX7Neo/ΔEX7Hygro (−/−).