Generation of mice containing a floxed SNAP-23 allele.

<p>(A) A 179 kb BAC clone containing the entire SNAP-23 gene was mutated by introducing loxP sites surrounding exon II alone (#1 floxed allele) or surrounding exons III-V (#2 floxed allele). Expression of Cre will result in the excision of exon II alone in the #1 floxed allele construct or exons III-V in #2 floxed allele construct. Each deleted allele introduces a frame shift that prevents the synthesis of in-frame SNAP-23 exon-encoded amino acids. (B) Typical PCR genotyping analysis of mice used in this study. The upper panel shows results obtained using a PCR reaction that can distinguish the deleted SNAP-23 allele present in SNAP-23^+/- mice (492 bp) from either the wild-type SNAP-23 allele or the floxed SNAP-23 BAC transgene (266 bp). The lower panel shows results obtained using a PCR reaction that can distinguish the floxed SNAP-23 BAC transgene (195 bp) from the wild-type SNAP-23 allele present in SNAP-23^+/- mice (161 bp). (C) Spleen cells were isolated from wild-type mice (SNAP-23^+/+) and two representative floxed SNAP-23 BAC transgene mice on a SNAP-23-/- background (SNAP-23^fl/-) and the cells were lysed and analyzed by SDS-PAGE and immunobloting with SNAP-23 and actin antibodies. Quantitative densitometry revealed that the floxed SNAP-23 BAC expression of SNAP-23 was approximately 50% that found in wild-type mice.