Fig. 1.

Generation of the GLYT2-Cre knock-in mouse line. (A) Schematic representations of the wild-type allele (GLYT2⁺), knock-in vector, targeted allele (GLYT2^Crefrtneo) and modified targeted allele after deletion of the PGK-neo-p(A) cassette (neo) (GLYT2^Cre). The black numbered boxes represent exons. The black triangle and the gray semicircles represent translation initiation sites and FRT sequences, respectively. The white boxes represent the Cre recombinase gene followed by polyA signal (Cre) and the neo cassette, respectively. The gray box and the line represent the MC1 promoter-driven diphtheria toxin A-fragment gene (DT-A) and the plasmid vector, respectively. The black circles represent the 5′ and 3′ termini of the homologous recombination region in the knock-in targeting vector. The red, green and blue boxes represent the positions of probes A, B and C, respectively. The expected restriction fragment lengths (in kb) for Southern blot analyses are shown. The small black arrows represent the positions of primers P1, P2, P3 and P4 for the PCR analysis. EI, EcoRI; EV, EcoRV. (B) Southern blot analysis of genomic DNA extracted from neomycin-resistant ES cell clones. B6 indicates the negative control. All twelve analyzed clones were correctly targeted. (C) PCR analysis of the progeny of GLYT2^Crefrtneo/+ mice crossed with FLP mice. GLYT2-Cre knock-in (GLYT2^Cre/+) mice and wild-type (GLYT2^+/+) mice were obtained. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)