Goat uromodulin promoter directs kidney-specific expression of GFP gene in transgenic mice.
BACKGROUND
Uromodulin is the most abundant protein found in the urine of mammals. In an effort to utilize the uromodulin promoter in order to target recombinant proteins in the urine of transgenic animals we have cloned a goat uromodulin gene promoter fragment (GUM promoter) and used it to drive expression of GFP in the kidney of transgenic mice.


RESULTS
The GUM-GFP cassette was constructed and transgenic mice were generated in order to study the promoter's tissue specificity, the GFP kidney specific expression and its subcellular distribution. Tissues collected from three GUM-GFP transgenic mouse lines, and analyzed for the presence of GFP by Western blotting and fluorescence confirmed that the GUM promoter drove expression of GFP specifically in the kidney. More specifically, by using immuno-histochemistry analysis of kidney sections, we demonstrated that GFP expression was co-localized, with endogenous uromodulin protein, in the epithelial cells of the thick ascending limbs (TAL) of Henle's loop and the early distal convoluted tubule in the kidney.


CONCLUSIONS
The goat uromodulin promoter is capable of driving recombinant protein expression in the kidney of transgenic mice. The goat promoter fragment cloned may be a useful tool in targeting proteins or oncogenes in the kidney of mammals.
Huang
Yue-Jin
YJ
PharmAthene Canada Inc, 1000 St-Charles Avenue Block B, Vaudreuil-Dorion, QC J7V 8P5, Canada. yjhuang@pharmathene.ca
Chretien
Nathalie
N
Bilodeau
Annie S
AS
Zhou
Jiang Feng
JF
Lazaris
Anthoula
A
Karatzas
Costas N
CN
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D005821:Genetic Techniques; D006041:Goats; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D007668:Kidney / Q000196:embryology* / Q000378:metabolism*; D007684:Kidney Tubules / Q000196:embryology / Q000378:metabolism; D008138:Loop of Henle / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D009088:Mucoproteins / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000737:chemistry; D013997:Time Factors; D014018:Tissue Distribution; D019076:Transgenes; D058949:Uromodulin
D009088:Mucoproteins; D011994:Recombinant Proteins; C548613:Umod protein, mouse; D058949:Uromodulin; D049452:Green Fluorescent Proteins