Design and testing of a targeting construct for the mouse Gimap6 gene.

<p>Panel A shows the targeting construct in which exons 2 and 3 are flanked by loxP sites, both before and after flpE recombination to remove the selectable neomycin resistance cassette (neo). In addition, DNA fragments encoding thymidine kinase (TK) and diphtheria toxin A chain (DTA) were included in the targeting construct to increase the targeting efficiency in embryonic stem cells. The lower part of panel A shows the integrated DNA fragment after ablation of exons 2 and 3 by Cre-mediated recombination. Panel B shows a Western blot of GIMAP6 (using rat anti-mouse antibody MAC436) and actin on total lymphocytes isolated from thymus or spleen of GIMAP6^fl/fl mice, in which the Gimap6 gene is intact, and in GIMAP6^fl/flCD2Cre mice, in which Gimap6 should be deleted in the B and T cell populations.