Tg(CLCNKB-WNK4)1Suc

An 11-kb human genomic DNA fragment that includes the promoter and first intron of the chloride channel Kb (CLCNKB) gene, extending from the 3' end of the CLCNKA gene to immediately upstream of the translation start site of CLCNKB, is joined to the 5' end of a cDNA encoding a C-terminal hemagglutinin (HA) tagged, mutant human WNK lysine deficient protein kinase 4 with alanine replacing aspartic acid at amino acid position 564 (D564A), which resides in a highly conserved segment distal to the first coil domain of the protein, and followed by an SV40 polyadenylation signal. Immunoblot analysis of kidney homogenate using anti-HA antibody demonstrated expression in the kidney. Immunohistochemical analysis localized kidney expression of the mutant protein to the distal nephrons of the cortex and outer medulla, the thick ascending limb of Henley's loop (TAL) and, variably, cells of the distal tubules. In the last, signal is localized to the cytoplasm, while in TAL it is concentrated at the apical membrane. (J:110994)