Kidney-specific activity of the bovine uromodulin promoter.
A 10-kilobase (kb) lambda bacteriophage bovine genomic clone containing 5.4 kb of the 5'-flanking region, exons, and introns of bovine uromodulin gene was isolated. Transgenic mice containing 3.9 kb of the bovine uromodulin promoter and a lacZ reporter gene were generated by pronuclear microinjection. RT-PCR and northern blot analyses of transgene expression in various tissues of founder and F1 mice showed that the transgene was expressed exclusively in the kidney. In situ hybridization and histochemistry for lacZ demonstrated that transgene expression was restricted to tubule epithelial cells of the loop of Henle in the kidney. Stepwise 5' deletion analysis revealed that transfection of luciferase reporter constructs fused to various proximal 5'-flanking regions of the bovine uromodulin gene markedly increased luciferase activity in mouse renal epithelial cells but not in mesenchymal cells and that the most critical cis elements of the uromodulin gene are located within the 600 bp upstream region.
In2Gen Co., Cancer Research Institute, Seoul National University College of Medicine, Yeongun-Dong 28, Jongro-Gu, Seoul 110-799, Republic of Korea. htkim@in2gen.com
Hun-Taek
HT
Kim
In-Young
IY
Song
Jorge
J
Piedrahita
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D017931:DNA Primers; D017403:In Situ Hybridization; D007684:Kidney Tubules / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D009088:Mucoproteins / Q000235:genetics*; D041681:NIH 3T3 Cells; D011401:Promoter Regions, Genetic; D058949:Uromodulin
D017931:DNA Primers; D009088:Mucoproteins; C548613:Umod protein, mouse; D058949:Uromodulin