Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
권호 표지
DOI QR코드

DOI QR Code

Molecular Cloning and mRNA Expression of the Hanwoo CAT/enhancer-binding Protein α(C/EBPα) Gene

한우 CCAAT/enhancer-binding protein α(C/EBPα) 유전자의 동정과 mRNA의 발현

  • Jeoung, Y.H. (Department of Animal Science, Insti. of Ag. Sci. and Tech. , College of Agriculture & Life Science, Chonnam National University) ;
  • Lee, S.M. (Department of Animal Science, Insti. of Ag. Sci. and Tech. , College of Agriculture & Life Science, Chonnam National University) ;
  • Park, H.Y. (Department of Animal Science, Insti. of Ag. Sci. and Tech. , College of Agriculture & Life Science, Chonnam National University) ;
  • Yoon, D.H. (National Livestock Research Institute, RDA) ;
  • Moon, S.J. (Department of Animal Science, Insti. of Ag. Sci. and Tech. , College of Agriculture & Life Science, Chonnam National University) ;
  • Chung, E.R. (Department of Biotechnology, College of Life Science and Natural Resources, Sangji University) ;
  • Kang, M.J. (Department of Animal Science, Insti. of Ag. Sci. and Tech. , College of Agriculture & Life Science, Chonnam National University)
  • 정영희 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 이상미 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 박효영 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 윤두학 (농촌진흥청 축산연구소) ;
  • 문승주 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 정의룡 (상지대학교) ;
  • 강만종 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부)
  • Published : 2004.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

CCAAT/enhancer binding proteins(C/EBP) are a group of transcription factors expressed during preadipocyte differentiation. In the C/EBPs, C/EBPa plays an important role in lipid deposition and adipocyte differentiation. In this studies, we report the identification, characterization, and expression of a Hanwoo CIEBP$\alpha$ The Hanwoo C/EBP$\alpha$DNA includes a 1059 bp open reading frame encoding a protein of 353 amino acids. The CIEBPa amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The distribution of C/EBP$\alpha$ mRNA in various tissues of Hanwoo aged 12 months were investigated using Northern blotting analysis. The highest expression was detected in adipose tissue and more lower expression was detected in colon and lung. We also identified expression of C/EBPa mRNA in Hanwoo sirloin and adipose tissue aged 12, 26, and 30 months by real-time RT-PCR. The higest expression were detected at 26 months in the sirloin and at 12 and 26 months in the adipose tissue.

CCAAT/enhancer binding protein(C/EBP)는 지방전구세포의 분화과정에서 말현하는 전사인자군에 속한다. 이러한 C/EBP 중에서 C/EBP $\alpha$ 는 지방세포의 분화와 지방침착에 있어 중요한 역할을 수행허는 것으로 알려져 있다. 본 연구에서는 한우 C/EBP$\alpha$ 유전자를 분리 동정하고 그 발현을 조사하였다. 한우 C/EBP$\alpha$ 유전자는 1059bp의 open reading frame으로 구성되어 있으며 353개의 아미노산을 암호화하고 있었다. 그리고 한우 C/EBP $\alpha$ 아미노산을 다른 종과 비교하였을 때 매우 높은 상동성을 나타내었다. Northern blotting 분석에 의하여 12개월 한우에 있어서 C/EBP$\alpha$ mRNA의 분석을 실시한 결과 지방조직에서 가장 높은 말현을 나타내었으며 대장과 폐에서 매우 낮은 말현을 확인할수 있었다. 또한 12개월26개월 30개월 한우 의 등심과 지방에 있어서 C/EBP$\alpha$ 유전자의 발현을 real-time RT-PCR로 분석한 결과 등심 에서는 26개월에 서 가장 높은 발현을 보였으며 지방에서는 12개월과 26개월에 서 높은 발현을 나타내었다.

Keywords

References

  1. Antonson, P. and Xanthopoulos, K. G. 1995. Molecular cloning, sequence, and expression patterns of the human gene encoding CCAAT/enhancer binding protein alpha(C/EBP alpha). Biochem. Biophys. Res. Commun. 215:106.
  2. Birkenmeier, E. H., Gwynn, B., Howard, S., Jerry, J., Gordon, J. I., Landschulz, W. H and McKnight, S. L. 1989. Tissue-specific expression, developmental regulation, and genetic mapping of the gene encoding CCAAT/enhancer binding protein. Genes Dev. 3:1146.
  3. Christy, R. J., Kaestner, K. H., Geiman, D. E. and Lane, M. D. 1991. CCAAT/enhancer binding protein gene promoter: binding of nuclear factors during differentiation of 3T3-L1 preadipocytes. Proc. Natl. Acad. Sci. USA 88:2593.
  4. Cao, Z., Umdk, R. M. and McKnight, S. L. 1991. Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells. Genes Dev. 5:1538.
  5. Cowherd, R M., Lyle, R E. and McGehee Jr, R. E. 1999. Molecular regulation of adipocyte differentiation. Semin. Cell. Dev. BioI. 10:3.
  6. Grimaldi, P. A. 2001. The roles of PPARs in adipocyte differentiation. Prog. Lipid. Res. 40:269.
  7. Ihara, N., Yamakuchi, H., Hirano, T., Takeda, H., Taniguchi, Y., Sasaki, Y., Davis, S. K., Taylor, J. F., Barendse, W. and Sugimoto, Y. 1997. Physical and genentic mapping of bovine CEBPA and PPARG genes. Anim. Genet. 29:398.
  8. Landschulz, W. H, Johnson, P. F., Adashi, E. Y., Graves, B. J. and McKnight, S. L. 1988a. Isolation of a recombinant copy of the gene encoding C/EBP. Genes Dev. 2:789.
  9. Landschulz, W. H., Johnson, P. F. and McKnight, S. L. 1988b. The lucine zipper: A hypothetical structure common to a new class of DNA binding proteins. Science. 240:1759.
  10. Manchado, C., Yubero, P., Vinas, O., Iglesias, R, Villarroya, F., Mampel, T. and Giralt, M. 1994. CCAAT/binding proteins a and $\beta$ in brown adipose tissue: evidence for a tissue-specific pattern of expression during development. Biochem. J. 302:695.
  11. Miller, S. A, Kykes, D. D. and Polesky, H. F. 1998. A simple salting out procedure for extracting DNA from human nucleated cells. Nucl. Acids. Reg. 16:1215.
  12. Taniguchi, Y. and Sasaki, Y. 1996. Rapid communication: Nucleotide sequence of bovine C/EBP $\alpha$ gene. J. Anim. Sci. 74:2554.
  13. Tchoukalova, Y. D., Hausman, D. B., Dean, R. G. and Hausman, G. J. 2000. Enhancing effect of troglitazone on porcine adipocyte differentiation in primary culture: A comparison with dexamethasone. Obes. Res. 8:83.
  14. Yu, Z. K. and Hausman, G. J. 1998. Expression of CCAAT/enhancer binding protein during porcine preadipocyte differentiation. Exp. Cell. Res. 245:343.