Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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Characterization of the Nanog 5'-flanking Region in Bovine

  • Choi, Don-Ho (The Institute of Hankook Life Science) ;
  • Kim, Duk-Jung (The Institute of Hankook Life Science) ;
  • Song, Ki-Duk (Department of Animal Biotechnology, Chonbuk National University) ;
  • Park, Hwan-Hee (Department of Biomedical Sciences, School of Medicine, Inha University) ;
  • Ko, Tae Hyun (Department of Biomedical Sciences, School of Medicine, Inha University) ;
  • Pyao, Yuliya (Department of Biomedical Sciences, School of Medicine, Inha University) ;
  • Chung, Ku-Min (The Institute of Hankook Life Science) ;
  • Cha, Seok Ho (Department of Parasitology and Tropical Medicine, School of Medicine, Inha University) ;
  • Sin, Young-Su (Department of Animal Science, Singu University) ;
  • Kim, Nam-Hyung (Department of Animal Science, Chungbuk National University) ;
  • Lee, Woon-Kyu (Department of Biomedical Sciences, School of Medicine, Inha University)
  • Received : 2016.01.12
  • Accepted : 2016.04.07
  • Published : 2016.10.01
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Abstract

Bovine embryonic stem cells have potential for use in research, such as transgenic cattle generation and the study of developmental gene regulation. The Nanog may play a critical role in maintenance of the undifferentiated state of embryonic stem cells in the bovine, as in murine and human. Nevertheless, efforts to study the bovine Nanog for pluripotency-maintaining factors have been insufficient. In this study, in order to understand the mechanisms of transcriptional regulation of the bovine Nanog, the 5'-flanking region of the Nanog was isolated from ear cells of Hanwoo. Results of transient transfection using a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the -134 to -19 region contained the positive regulatory sequences for the transcription of the bovine Nanog. Results from mutagenesis studies demonstrated that the Sp1-binding site that is located in the proximal promoter region plays an important role in transcriptional activity of the bovine Nanog promoter. The electrophoretic mobility shift assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. In addition, significant inhibition of Nanog promoter activity by the Sp1 mutant was observed in murine embryonic stem cells. Furthermore, chromatin-immunoprecipitation assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. These results suggest that Sp1 is an essential regulatory factor for bovine Nanog transcriptional activity.

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