Expression and regulation of hFIX minigene and cDNA driven by β-casein gene in mouse mammary gland |
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Authors: | ZHANG KezhongJIANG PeihongLU DaruHUANG WeidaCHEN LiXUE Jerry L. HsuehQIU Xinfang |
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Affiliation: | (1) Institute of Genetics, Fudan University, 200433 Shanghai, China;(2) Department of Biochemistry, Fudan University, 200433 Shanghai, China |
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Abstract: | Mammary gland specific expression vectors for human clotting factor IX (hFIX) and LacZ reporter gene driven by bovine β-casein gene were constructed. Vectors were packaged by stearylamine (SA) liposome and were transferred to lactating mice via tail vein. Both hFIX and Lac2 gene could be expressed in the mammary gland of the treated mice. The highest production of hFIX protein was 80.28 ng per mL milk, and more than 85% of hFIX protein appeared to be γ-carboxylation and biologically active. The results suggested that the 2.0 kb sequence of β-casein gene including promoter, exon 1 was effective to drive hFIX gene expression in mammary gland and intron 1 of β-casein gene had an effect on the tissue specific expression. The expression level in mouse milk injected with hFIX minigene vector containing hFIX endogenous intron 1 was increased by above 3 times of that injected with hFIX cDNA vector. Project supported by the State High Technology Development Program and Shanghai Science and Technology Commission. |
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Keywords: | β -casein gene human clotting factor IX (hFIX) mammary gland expression |
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