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Effects of the cis-Acting Element in the 3' End of Porcine $\beta$-Casein Gene on the Expression in Mammary Epithelial Cells  

Lee, Hwi-Cheul (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Kim, Byoung-Ju (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Byun, Sung-June (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Lee, Seung-Hoon (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Kim, Min-Ji (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Chung, Hee Kyoung (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Lee, Hyun-Gi (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Jo, Su-Jin (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Chang, Won-Kyong (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Park, Jin-Ki (Animal Biotechnology Division, National Livestock Research Institute, RDA)
Lee, Poong-Yeon (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Publication Information
Reproductive and Developmental Biology / v.32, no.3, 2008 , pp. 153-158 More about this Journal
Abstract
Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.
Keywords
mRNA stability; Porcine $\beta$-casein gene; Luciferase assay;
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