FIG. 2.

Targeted disruption of the TACC2 gene. (A) Schematic structure of and targeting strategy for the murine TACC2 locus. Empty boxes represent exons 4 to 12 of the murine TACC2 gene. The locations of the 5′ external probe and PCR primers for genotyping are indicated. Restriction enzyme sites are as follows: E, EcoRI; A, AflII; X, XbaI. wt, wild type; NEO, neomycin resistance cassette; DTA, diphtheria toxin A cassette; ko, knockout. Confirmation of successful targeting of ES cells was obtained by Southern blot analysis (B) and PCR screening of F₁ mice for the presence of the disrupted allele (C). WT, wild type; KO, knockout; HET, heterozygote. (D) Northern blot analysis of kidney tissue with a TACC2-specific probe shows no hybridization signal in knockout tissue. Note that disruption of the TACC2 gene led to the complete absence of both the 4- and 10-kb transcripts, therefore generating a null mutation for all transcript variants. Hybridization for actin (lower panel) was used to control RNA loading. The positions of transcript sizes are indicated.