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Characterization of bovine 20a‐HSD gene promoter
Author(s) -
Min KwanSik,
Purevjargal Naidansuren,
Kang MyeongHwa
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.906.2
Subject(s) - microbiology and biotechnology , tata box , transfection , gene , promoter , biology , expression vector , mutant , vector (molecular biology) , corpus luteum , reporter gene , gene expression , genetics , recombinant dna , ovary
The 20¥á‐hydroxysteroid dehydrogenase (20¥á‐HSD) enzyme catalyzes the NADPH‐dependent reduction of progesterone to 20¥á‐hydroxypregn‐4‐ene‐3‐ one (20¥á‐hydroxyprogesterone), which is biologically inactive. Thus, it is very important enzyme to maintenance of pregnancy in mammlian. We cloned the promoter (−927bp) of bovine 20a‐HSD gene to charaterize the functional role and constructed five deleted mutants into the pGL3‐basic vector. And we also constructed the EGFP gene vector under bovine 20a‐HSD promoter. Bovine 20a‐HSD gene promoter contains TATA box, two putative CCAAT boxes and several transcriptional factor sites (GATA‐1, CRE‐BP, Oct‐1, Ap‐1 and SRY). The promoter activity was remarkably decreased in all deleted vector using the rat corpus luteum cells. But, it was recovery by the co‐transfection of the Sp‐1 factor. The constructed vector (promoter+EGFP) was expressed EGFP signal into the CHO K1 cell at 2 days after transfection. (This work was supported by a grant (20080401034074) from BioGreen 21 Program, Rural Development Administration, Republic of Korea)