Klrk1^tm1.1Bpol

The locus contains two alternative start sites, one in exon 2 and one in exon 3. The ATG start site in exon 2 was mutated into an EcoRI restriction site. Exon 3 was replaced with a cassette containing an EGFP coding sequence, a transcriptional stop signal, and a loxP-flanked neomycin resistance cassette. Correctly targeted ES cells had the neomycin cassette removed by transient expression of cre recombinase. Heterozygote natural killer (NK) cells had reduced protein expression of NKG2D while no protein expression was detected on homozygote NK cells as determined by flow cytometry. EGFP expression was not detected and subsequent sequence analysis of the targeting vector revealed the Kozak sequence at the leading ATG of the EGFP coding sequence was deleted during sub-cloning. (J:151878, J:215590)