title: Ovarian and placental expression of 20α-hydroxysteroid dehydrogenase during pregnancy in deer.
abstract: The enzyme 20α-hydroxysteroid dehydrogenase (20α-HSD) catalyzes the conversion of progesterone to its inactive form, 20α-hydroxyprogesterone. This enzyme has been shown to play a critical role in the regulation of luteal function in experimental animals. In this study, we cloned and expressed the gene encoding elk deer 20α-HSD from reproductive placental and ovarian tissues. PCR, 3'- and 5'-RACE, and northern blot analysis were performed for the cloning and characterization of deer 20α-HSD gene. We expressed recombinant deer 20α-HSD protein and used western blot analysis to determine protein expression levels in the placenta and ovary during pregnancy. The full cDNA sequence of 20α-HSD was used to clone an open reading frame encoding 323 amino acids and consisting of 1142 bp. The nucleotide sequence of deer 20α-HSD showed high homology with the sequences of the bovine (96%), goat (96%), and human (83%) 20α-HSD genes. 20α-HSD mRNA was strongly expressed in the placenta on days 30, 60, and 70 of pregnancy. A high level of the protein was also detected in the placenta but not in fetal skin tissue. The recombinant 20α-HSD protein produced in mammalian cells and bacterial systems had a molecular weight of approximately 37-kDa. The deer 20α-HSD protein signal was specifically localized in the basal part of the primary chorionic villi and chorionic stem villus of the placenta during early pregnancy. The 20α-HSD protein was also intensively localized in the larger luteal cells of the corpus luteum during pregnancy.
authors.affiliation: Animal Biotechnology, Graduate School of Bio & Information Technology, Institute of Genetic Engineering, Hankyong National University, Ansung, Republic of Korea.
authors.forename: P J
authors.initials: PJ
authors.lastname: Naidansuren
authors.forename: C W
authors.initials: CW
authors.lastname: Park
authors.forename: T M
authors.initials: TM
authors.lastname: Nanjidsuren
authors.forename: J J
authors.initials: JJ
authors.lastname: Park
authors.forename: S J
authors.initials: SJ
authors.lastname: Yun
authors.forename: M H
authors.initials: MH
authors.lastname: Kang
authors.forename: K
authors.initials: K
authors.lastname: Yamanouchi
authors.forename: K S
authors.initials: KS
authors.lastname: Min
mesh_terms: D015089:20-Hydroxysteroid Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D003670:Deer; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D008969:Molecular Sequence Data; D010053:Ovary / Q000378:metabolism*; D010920:Placenta / Q000502:physiology*; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D015089:20-Hydroxysteroid Dehydrogenases