Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Construction and Verification of Useful Vectors for Ectopic Expression and Suppression of Plant Genes.

식물 유전자의 과발현 및 발현 억제를 위한 유용 벡터의 제조 및 확인

  • Lee, Young-Mi (Department of Molecular Biology, Pusan National University) ;
  • Seok, Hye-Yeon (Department of Molecular Biology, Pusan National University) ;
  • Park, Hee-Yeon (Department of Molecular Biology, Pusan National University) ;
  • Park, Ji-Im (Department of Molecular Biology, Pusan National University) ;
  • Han, Ji-Sung (Department of Molecular Biology, Pusan National University) ;
  • Bang, Tae-Sik (Department of Molecular Biology, Pusan National University) ;
  • Moon, Yong-Hwan (Department of Molecular Biology, Pusan National University)
  • 이영미 (부산대학교 분자생물학과) ;
  • 석혜연 (부산대학교 분자생물학과) ;
  • 박희연 (부산대학교 분자생물학과) ;
  • 박지임 (부산대학교 분자생물학과) ;
  • 한지성 (부산대학교 분자생물학과) ;
  • 방태식 (부산대학교 분자생물학과) ;
  • 문용환 (부산대학교 분자생물학과)
  • Published : 2009.06.30
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Abstract

The phenotypes associated with a gene function are often the best clue to its role in the plant. Transgenic plants ectopically expressing or suppressing a gene can provide useful information related to the gene function. In this study, we constructed three vectors - pFGL571, pFGL846 and pFGL847 - for the Agrobacterium-mediated ectopic expression of plant genes using pPZP211 and modified CaMV 35S, UBQ3 or UBQ10 promoters. The three vectors have several merits such as small size, high copy in bacteria, enough restriction enzyme sites in multi cloning sites and nucleotide sequence information. Analysis of transgenic plants containing GUS or sGFP reporter genes under the control of modified CaMV 35S, UBQ3 or UBQI0 promoter revealed that all of the three promoters showed high activities during most developmental stages after germination and in floral organs. Furthermore, we generated a RNAi module vector, pFGL727, to suppress plant gene expressions and confirmed that pFGL727 is useful for the suppression of a gene expression using rice transgenic plants. Taken together, our new vectors would be very useful for the ectopic expression or the suppression of plant genes.

식물에서 유전자의 기능을 연구하는데 있어서 유전자가 과발현 되거나 발현이 억제되는 형질전환체는 해당 유전자의 기능과 관련되어 매우 유용한 정보를 제공한다. 본 연구에서는 modified CaMV 355, UBQ3, UBQ10 프로모터를 pPZP211 벡터에 각각 클로닝 하여 Agrobacterium을 매개로 한 과발현형질전환 식물체 제작에 유용하게 이용할 수 있는 pFGL571, pFGL846, pFGL847을 제조하였다. 이 벡터들은 크기가 작고, 박테리아 내에 high copy로 존재하며, 다중 클로닝 부위에 다양한 제한효소 부위를 가지고 있고, 전체 서열이 알려져 있는 등의 장점을 가지고 있다. GUS 또는 sGFP 리포터 유전자를 포함하는 형질전환 식물체를 제조하여 modified CaMV 35S, UBQ3, UBQ10 프로모터의 활성을 분석한 결과, 세 프로모터 모두 발아 후 대부분의 발달단계와 성숙한 식물체의 꽃 기관에서 높은 활성을 보였다. 한편, 식물에서 유전자 발현 억제에 이용할 수 있는 RNAi 기본 벡터인 pFGL727을 제조하였고, pFGL727을 이용한 벼 RNAi 형질전환체의 분석을 통해 이벡터가 유전자의 발현 억제에 유용하게 이용될 수 있음을 확인하였다. 연구 결과를 종합해 보면, 본 연구에서 제조한 벡터들은 식물에서 유전자 과발현과 발현 억제에 유용하게 이용될수 있을 것으로 기대된다.

Keywords

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