Tg(Ube3a)2Bdph

The bacterial artificial chromosome (BAC) clone RP24-178G7 contains the entire ubiquitin protein ligase E3A locus (Ube3a) as well as ~63 kbp of 5' flanking sequence and ~21 kbp of 3' flanking sequence. DNA sequences encoding a floxed Ube3a exon 7 (as per Ube3atm1.1Bdph) were introduced to the BAC via homologous recombination/BAC recombineering, replacing the native Ube3a exon 7 and surrounding sequences and resulting in a conditional transgenic allele. A single frt site remains just upstream of floxed Ube3a exon 7. The transgenic insertion in the founder line was determined to comprise two tandem copies of the Tg(Ube3a)1Bdph transgene. Crossing to FLPo delete mice resulted in efficient recombination of the FRT sites, enabling the production of a subline of mice with a single Tg(Ube3a)1Bdph copy. Ube3a expression was confirmed by qPCR, Western blot analysis, and immunofluorescent staining. (J:302578)