Goat uromodulin promoter directs kidney-specific expression of GFP gene in transgenic mice

被引:8
作者
Huang, YJ [1 ]
Chretien, N [1 ]
Bilodeau, AS [1 ]
Zhou, JF [1 ]
Lazaris, A [1 ]
Karatzas, CN [1 ]
机构
[1] PharmAthene Canada Inc, Vaudreuil Dorion, PQ J7V 8P5, Canada
关键词
D O I
10.1186/1472-6750-5-9
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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. Conclusion: 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.
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页数:13
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