Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Biological Activity of Recombinant Human Granulocyte Colony-Stimulating Factor and Isolation of the Somatic Cell Transfected EGFP-hG-CSF Gene

유전자 재조합 인간의 G-CSF의 생리활성과 EGFP-hG-CSF유전자가 도입된 체세포의 분리

  • Park, Jong-Ju (Animal Biotechnology, The Graduate School of Bio. & Information Technology, Institute of Genetic Engineering, Hankyong National University) ;
  • Min, Kwan-Sik (Animal Biotechnology, The Graduate School of Bio. & Information Technology, Institute of Genetic Engineering, Hankyong National University)
  • 박종주 (한경대학교 생물환경정보통신전문대학원, 유전공학연구소, 동물생명공학) ;
  • 민관식 (한경대학교 생물환경정보통신전문대학원, 유전공학연구소, 동물생명공학)
  • Published : 2008.07.30
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Abstract

To investigate the biological activity of recombinant human granulocyte colony-stimulating factor (rec-hG-CSF) in mammalian cells, hG-CSF gene was cloned using the eDNA extracted from the human squamous carcinoma cell lines and rec-hG-CSF was produced in CHO cell lines. To analyze the biological activity in vivo, the rec-hG-CSF protein was injected into mice subcutaneously on days 0 and 2. Blood was withdrawn for white blood cell (WBC) determination 5 days after the first injection. WBC values were found to have increased significantly. A pEGFP-mUII-hG-CSF vector was transfected into somatic cell lines isolated from bovine fetal cells. The colony expressing EGFP signals was observed with a confocal microscope. These data suggest that the rec-hG-CSF produced in this study has potent activity in vivo. Thus, the results of this biological activity show that rec-hG-CSF can be enhanced considerably by genetic engineering that affects potential activity, including mutations, which add the oligosaccharide chain and construct double-fusion proteins. A pEGFP-mUII-hG-CSF vector can be utilized for the production of cloned transgenic livestock.

유전자재조합 hG-CSF의 생리활성을 분석하기 위하여 편상의 암세포로부터 분리되어진 cDNA를 이용하여 hG-CSF 유전자를 분리하여 동물세포(CHO cell lines)를 이용하여 재조합 단백질을 생산하였다. 재조합 단백질의 체내 생리활성을 분석하기 위하여0일과 2일에 피하주사 후 5일에 혈액을 채취하여 백혈구 수를 분석하였다. 투여 전과 비교하여 5일째에 백혈구 수는 현저하게 증가하였다. 또한, pEGFP-mUII-hG-CSF벡터를 소 태아로부터 분리되어진 체세포에 형질전환을 시켜서, EGFP signal을 나타내는 세포를 confocal를 이용하여 분리하여 수립하였다. 따라서, 이러한 결과는 유전자재조합 hG-CSF는 체내에서 강력한 생리활성을 나타내며, 또한 당쇄가 첨가되어지고 이중으로 연결되어진 새로운 돌연변이체를 포함하여 고 활성 재조합체의 생산이 가능할 것으로 보이며, pEGFP-mUII-hG-CSF벡터는 복제 형질전환 가축 생산을 위하여 유용하게 사용되어질 것으로 사료된다.

Keywords

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