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Effect of IRES Controlled Reporter Gene on Screening and Production of Recombinant Human EPO Proteins from Cultured CHO Cells

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  • 발행기관
    한국동물생명공학회(구 한국동물번식학회) 바로가기
  • 간행물
    Reproductive & developmental biology 바로가기
  • 통권
    Volume 30 No 2 (2006.06)바로가기
  • 페이지
    pp.81-85
  • 저자
    Hyun Gi Lee, Jin-Ki Park, Sung-Woo Kim, Eun-Mi Ko, Byoung-Ju Kim, Su-Jin Jo, Sung-June Byun, Boh-Suk Yang, Won-Kyong Chang, Hoon Taek Lee, Poongyeon Lee
  • 언어
    영어(ENG)
  • URL
    https://www.earticle.net/Article/A36252

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초록

영어
This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA. Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.

목차

INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

키워드

hEPO Glycosylation site Site-directed mutagenesis IRES GFP CHO cell

저자

  • Hyun Gi Lee [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]
  • Jin-Ki Park [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]
  • Sung-Woo Kim [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]
  • Eun-Mi Ko [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]
  • Byoung-Ju Kim [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]
  • Su-Jin Jo [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]
  • Sung-June Byun [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]
  • Boh-Suk Yang [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]
  • Won-Kyong Chang [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]
  • Hoon Taek Lee [ Department of animal Biotechnology, KonKuk University ]
  • Poongyeon Lee [ Transgenic Research Lab, Animal Biotechnology Division, NLRI, RDA ]

참고문헌

자료제공 : 네이버학술정보

간행물 정보

발행기관

  • 발행기관명
    한국동물생명공학회(구 한국동물번식학회) [The Korean Society of Animal Reproduction and Biotechnology]
  • 설립연도
    1976
  • 분야
    농수해양>축산학
  • 소개
    동물번식생리학, 동물생명공학, 수의학, 인공수정 및 수정란이식을 이용한 동물개량에 관한 이론과 기술의 발전을 통해 학계, 연구계, 산업계 및 양축가 상호간의 협력을 도모함으로써 동물과학발전 및 사회일반의 이익에 기여 한다는 목적을 위해 노력해 나가겠습니다.

간행물

  • 간행물명
    Reproductive & developmental biology
  • 간기
    계간
  • pISSN
    1738-2432
  • eISSN
    2288-0151
  • 수록기간
    1977~2018
  • 십진분류
    KDC 527 DDC 636

이 권호 내 다른 논문 / Reproductive & developmental biology Volume 30 No 2

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