Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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A Novel Oxidative Stress-inducible Peroxidase Promoter and Its Applications to Production of Pharmaceutical Proteins in Transgenic Cell Cultures

  • Lee, Ok-Sun (Lab. of Plant Cell Biotechnology, Korea Research institute of Bioscience and Biotechnology(KRIBB)) ;
  • Park, Sun-Mi (Lab. of Environmental Biotechnology, Korea Research Institute of Bioscience and Biotechnology(KRIBB)) ;
  • Kwon, Suk-Yoon (Lab. of Environmental Biotechnology, Korea Research Institute of Bioscience and Biotechnology(KRIBB)) ;
  • Lee, Haeng-Soon (Lab. of Plant Cell Biotechnology, Korea Research institute of Bioscience and Biotechnology(KRIBB)) ;
  • Kim, Kee-Yeun (Lab. of Environmental Biotechnology, Korea Research Institute of Bioscience and Biotechnology(KRIBB)) ;
  • Kim, Jae-Whune (R & D Center, Microplants Co., LTD.) ;
  • Kwak, Sang-Soo (Lab. of Environmental Biotechnology, Korea Research Institute of Bioscience and Biotechnology(KRIBB))
  • Published : 2002.12.01
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Abstract

A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

Keywords

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