Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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In Vitro Expression of the Recombinant hFSH Gene using Retrovirus Vector System

In Vitro에서 Retrovirus Vector System을 이용한 재조합 hFSH 유전자의 발현

  • Min, Gyeong-Heon (Department of Physiology, Catholic University of Daegu School of Medicine) ;
  • Kwon, Mo-Sun (Department of Physiology, Catholic University of Daegu School of Medicine) ;
  • Kim, Teoan (Department of Physiology, Catholic University of Daegu School of Medicine) ;
  • Koo, Bon-Chul (Department of Physiology, Catholic University of Daegu School of Medicine)
  • 민경헌 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 권모선 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 김태완 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 구본철 (대구가톨릭대학교 의과대학 생리학교실)
  • Received : 2011.03.02
  • Accepted : 2011.03.07
  • Published : 2011.03.31
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Abstract

hFSH is a glycoprotein secreted from anterior pituitary and consists of ${\alpha}$ and ${\beta}$ subunits. Because of its major biological functions including sperm formation in the male and for follicular growth, FSH is used to cure woman's sterility. In this study we tried to produce recombinant hFSH in vitro using a retrovirus expression vector. Two major components of the vector we constructed are: ( i ) a DNA fragment containing ${\alpha}$ and ${\beta}$ genes fused by a DNA sequence coding carboxyl terminal peptide (CTP) of human chorionic gonadotropin, (ii) a DNA fragment corresponding woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). Evaluation of expression profile of the recombinant FSH using reverse transcription PCR and enzyme-linked immunosorbent assay (ELISA). Among three cell lines tested, HeLa cells were the best for hFSH expression (5,395 mIU/ml), then followed by chicken embryonic fibroblast (CEF) cells and Chinese hamster ovary (CHO) cells in the order of hFSH production. In addition to the amount, the FSH produced from HeLa cells was highest in terms of biological activity which was determined by measuring cAMP.

Keywords

References

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