Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Production of Green Fluorescent Protein (GFP) from Transgenic Rice Cell Suspension Culture

형질전환된 벼세포배양에서 green fluorescent protein (GFP) 생산

  • Lee, Jae-Hwa (Department of Pharmaceutical Engineering, College of Medical Life Science, Silla University)
  • 이재화 (신라대학교 의생명과학대 제약공학과)
  • Published : 2007.02.28
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Abstract

Green fluorescent protein (GFP) is an attractive reporter for bioprocess monitoring. A fluorescence-based method was developed to quantify GFP levels in transgenic plants and protein extracts. In this study, GFP was produced and secreted from suspension cells derived from transgenic rice. The RAmy3E promoter placed before the GFP gene controlled by sugars such as sucrose. The effects of sucrose concentration on the secretion of GFP and total protein into the medium were investigated in batch suspension culture. It was possible, therefore, to induce the expression of the GFP by removing sucrose from the cultured media or by allowing the rice suspension cells to deplete sucrose catabolically. The dry cell weight (7.06 g/L) and GFP level were detected as highest at 12%, 3% sucrose after 20 day culture, respectively. However secreted GFP fluorescence at the other sucrose concentrations (6%, 12%, 18% and 24%) were a little amount in media.

형광단백질 (green fluorescent protein, GFP)은 생물공정을 살피데 지표 단백질로 유용하게 사용이 된다. 본 연구에서는 쌀세포에서 외래 단백질의 발현양상을 관찰하기 위해서, 표지 단백질로 GFP를 형질전환 후 이것에서 유도된 현탁세포에서 GFP의 발현 양상을 관찰하였다. 형질전환시 GFP의 발현을 위한 프로모터로 RAmysE를 사용하였으며 이것은 배양액 중에서 당이 고갈되었을 때 강력히 작동된다. 그래서 본 연구에서는 배양액 중에 다양한 슈크로오스 농도로 쌀세포를 배양하여 세포의 성장양태 및 GFP의 발현양에 미치는 영향을 관찰한 결과 세포의 성장은 12%의 당농도에서 7.06g/L로 최적이였으며 GFP는 당을 가장 적게 사용한 3%에서 최적임을 알 수가 있었다. 이것은 세포의 성장과 GFP의 생산에 사용된 당이 반대로 영향을 미침을 알 수가 있었으며 향후 최적의 대량배양을 위해서는 세포의 성장과 산물의 생산시기를 분리한 이단계 배양법이 필요함을 암시한다.

Keywords

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