Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Transgenic Siberian Ginseng Cultured Cells That Produce High Levels of Human Lactoferrin

인체 락토페린 생산 형질전환 가시오갈피 배양세포

  • Jo Seung-Hyun (Laboratory of Plant Cell Biotechnology, Korea Research Institute of Bioscience and Biotechnology (KIRBB), Department Food Science and Technology, Chungnam National University) ;
  • Kwon Suk-Yoon (Laboratory of Environmental Biotechnology, Korea Research Institute of Bioscience and Biotechnology (KIRBB)) ;
  • Kim Jae-Whune (Microplants Co., LTD) ;
  • Lee Ki-Teak (Department Food Science and Technology, Chungnam National University) ;
  • Kwak Sang-Soo (Laboratory of Environmental Biotechnology, Korea Research Institute of Bioscience and Biotechnology (KIRBB)) ;
  • Lee Haeng-Soon (Laboratory of Plant Cell Biotechnology, Korea Research Institute of Bioscience and Biotechnology (KIRBB))
  • 조승현 (한국생명공학연구원 식물세포공학연구실, 충남대학교 식품공학과) ;
  • 권석윤 (한국생명공학연구원 환경생명공학연구실) ;
  • 김재훈 ((주)마이크로프랜츠) ;
  • 이기택 (충남대학교 식품공학과) ;
  • 곽상수 (한국생명공학연구원 환경생명공학연구실) ;
  • 이행순 (한국생명공학연구원 식물세포공학연구실)
  • Published : 2005.09.01
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Abstract

Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

락토페린은 철 결합 당단백질로서 항 미생물활성과 면역강화와 같은 생리활성 기능을 가지고 있다. 본 연구는 배양 세포 고 발현 SWPA2 promoter를 이용하여 인체락토페린(hLf)을 생산하는 형질전환 가시오갈피 배양세포주 개발에 관한 것이다. 형질전환에 이용된 벡터는 산화스트레스 유도성 SWPA2 promoter의 조절 하에서 hLf이 소포체로 targeting되도록 제작된 SWPA2pro::ER-hLf/pCAMBIA이다. hLf을 생산하는 각 형질전환 배양세포들은 PCR과 Southern분석을 통해 hLf 유전자가 가시오갈피 게놈내로 성공적으로 도입되었음을 확인하였으며, western blot과 ELISA를 통해 형질전환 가시오갈피 배양 세포주에서 hLf 단백질이 활성이 있음을 확인하였다. 형질전환 가시오갈피 배양세포에서 hLf 단백질의 함량은 세포배양이 진행될수록 증가하여 정지기 때 가장 높았으며 전체 수용성 단백질의 약 3%를 차지하였다. 따라서 본 연구에서 개발된 인체락토페린을 고생산하는 약용식물 가시오갈피 배양 세포주는 산업적으로 이용될 수 있을 것으로 기대된다.

Keywords

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  1. High-yield production of functional human lactoferrin in transgenic cell cultures of siberian ginseng (Acanthopanax senticosus) vol.11, pp.5, 2006, https://doi.org/10.1007/BF02932312