FIG. 1.

Generation and analysis of Kif3C^typeI and Kif3C^typeII mutants in ES cells. (A) Strategy for generating Kif3C^typeI and Kif3C^typeII mutants. A 2.1-kb DNA fragment between BamHI and EcoRV of the Kif3C gene was flanked with two loxP fragments. A PGK-neo and PGK-tk selection marker with a third loxP fragment was added into the vector. (B) Southern analysis of targeted ES cells (Kif3C^typeIII). ES cell genomic DNA was cut with BamHI and probed with a 600-bp NdeI DNA fragment (probe A). The wild-type and targeted Kif3C^typeIII alleles were detected as 9- and 26-kb bands, respectively. (C) Southern analysis of Cre-transfected ES cells. Kif3C^typeIII (+/−) ES cells were transfected with the Cre plasmid to excise the DNA fragment flanked by loxP sites. ES clones resistant to ganciclovir were analyzed by Southern analysis, in which the genomic DNA was digested with EcoRI and probed with a loxP fragment (probe B). Each band represents a loxP fragment. Two types (Kif3C^typeI and Kif3C^typeII) of ES cells were obtained.