Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Production of stable chloroplast-transformed plants in potato (Solanum tuberosum L.)

안정적 감자 엽록체 형질전환 식물체 생산

  • 민성란 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 정원중 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 박지현 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 유재일 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 이정희 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 오광훈 (한국생명공학연구원 식물시스템공학연구센터) ;
  • 정화지 ((주)젠닥스) ;
  • 유장렬 (한국생명공학연구원 식물시스템공학연구센터)
  • Received : 2011.03.04
  • Accepted : 2011.03.15
  • Published : 2011.03.31
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Abstract

Chloroplast genetic engineering of higher plants offers several unique advantages compared with nuclear genome transformation, such as high levels of transgene expression, a lack of position effect due to site-specific transgene integration by homologous recombination, multigene engineering in a single transformation event and reducing risks of gene flow via pollen due to maternal inheritance. We established a reproducible chloroplast transformation system of potato using a tobacco specific plastid transformation vector, pCtVG (trnI-Prrn-aadA-mgfp-TpsbA-trnA). Stable transgene integration into chloroplast genomes and the homoplasmic state of the transgenome were confirmed by PCR and Southern blot analyses. Northern, immunoblot analysis, and GFP fluorescence imaging revealed high expression and accumulation of GFP in the plastids of potato leaves. This system would provide new opportunities for genetic improvement and mass production of value added foreign proteins in this crop.

고등식물의 엽록체 유전공학은 핵 형질전환과 비교해 볼때 여러 가지 독특한 장점을 가진다. 높은 transgene 발현율, 상동 재조합에 의한 site-specific transgene의 삽입으로 인해 유전자의 position effect가 없으며, 단일 형질전환으로 동시에 여러 유전자의 도입이 가능하고 모계 유전으로 인해 화분 방출 위험을 감소시킬 수 있다. 담배 specific한 pCtVG (trnI-Prrn-aadA-mgfp-TpsbA-trnA) 벡터를 이용하여 안정적인 감자 엽록체 형질전환 시스템을 개발하였다. 감자 엽록체 게놈으로 외래유전자의 삽입과 homoplasmic level은 PCR과 Southern blot 분석으로 확인하였다. Northern과 immunoblot 분석 및 GFP fluorescence imaging을 통하여 엽록체 형질전환체의 잎에서 GFP 유전자가 강하게 발현, 축적됨을 알 수 있었다. 본 연구에서 확립된 감자 엽록체 형질전환 시스템을 이용하여 유용 유전자를 도입함으로써 농업적 형질을 개선하거나 고부가가치 단백질을 대량 생산하는 감자를 보다 효율적으로 개발할 수 있을 것이다.

Keywords

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