Target disruption of CIBZ in mouse ESCs.
A, WT genomic locus of CIBZ, targeting vector and the recombinant CIBZ locus. Bold box in the genomic locus represents the exon of the CIBZ gene. The open boxes in the targeting vector schematics represent pgk-tk and pgk-neo selectable marker genes. B, BamHI. B, Southern blot analysis of BamHI-digested genomic DNA from RF8 ESCs stably transfected with the targeting vector. DNA probe is indicated in A. C, PCR analysis of the recombinant locus using primers indicated by A (arrowheads). A 2.2 kb band results from the recombinant locus. D, generation of two CIBZ−/− ESC (CIBZ−/−1 and CIBZ−/−2) clones. CIBZ−/− homozygous cell lines were derived by selection using a high concentration of G418. Homologous recombination was confirmed by PCR using primers indicated in A. E, Western blotting showing CIBZ expression. α-Tubulin was used as a control for loading.