Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Construction and Analysis of Binary Vectors for Co-Overexpression, Tissue- or Development-Specific Expression and Stress-Inducible Expression in Plant

식물에서 표적 유전자의 동시 과발현, 조직/발달 특이적 발현 및 스트레스 유도성 발현을 위한 binary 벡터의 제작과 분석

  • Lee, Young-Mi (Department of Molecular Biology, Pusan National University) ;
  • Park, Hee-Yeon (Department of Molecular Biology, Pusan National University) ;
  • Woo, Dong-Hyuk (Department of Molecular Biology, Pusan National University) ;
  • Seok, Hye-Yeon (Department of Molecular Biology, Pusan National University) ;
  • Lee, Sun-Young (Ulsan University Hospital Biomedical Research Center, University of Ulsan) ;
  • Moon, Yong-Hwan (Department of Molecular Biology, Pusan National University)
  • 이영미 (부산대학교 분자생물학과) ;
  • 박희연 (부산대학교 분자생물학과) ;
  • 우동혁 (부산대학교 분자생물학과) ;
  • 석혜연 (부산대학교 분자생물학과) ;
  • 이선영 (울산대학교 의과대학 생의과학연구소) ;
  • 문용환 (부산대학교 분자생물학과)
  • Received : 2010.07.26
  • Accepted : 2010.09.07
  • Published : 2010.09.30
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Abstract

In this study, we constructed various kinds of binary vectors with the pPZP backbone for co-overexpression, tissue- or development-specific expression and stress-inducible expression, and validated them for ectopic expression of target genes. Using a modified CaMV 35S promoter, a binary vector was generated for co-overexpression of two different genes and was confirmed to be efficient for overexpressing two different target genes at the same time and place. Binary vectors containing At2S3, KNAT1 or LFY promoters were constructed for tissue-specific or development-specific gene expression, and the binary vectors were suited for embryo/young seedling stage-, shoot apical meristem- or leaf primordia-specific expressions. Furthermore, the binary vectors containing RD29A or AtNCED3 promoters were validated as suitable vectors for gene expression induced by abiotic stresses such as high salt, ABA, MV and low temperature. Taken together, the binary vectors constructed in this study would be very useful for analyzing the biological functions of target genes and molecular mechanisms through ectopic expression.

유전자를 이소성으로 발현하고 억제하는 것은 유전자의 기능 연구에 있어서 매우 유용하다. 본 연구에서는 표적 유전자의 동시 과발현, 조직/발달 단계 특이적 발현 및 스트레스 유도성 발현을 위해 pPZP를 골격으로 다양한 binary 벡터를 제작하고 그 유용성을 검증하였다. 변형된 CaMV 35S 프로모터를 이용하여, 다른 두 개의 유전자를 동시 과발현시키는 binary 벡터를 제작하였고, 이 벡터가 동시에 그리고 같은 장소에서 다른 두 개의 표적 유전자를 과발현 하는데 효과적임을 확인하였다. At2S3, KNAT1 및 LFY 프로모터를 포함하는 조직 또는 발달 단계 특이적 발현 binary 벡터들을 제작하고 분석한 결과, 이 벡터들은 각각 배/유식물 시기, 새싹 끝의 분열조직 및 잎 원기 특이적 발현에 유용하였다. RD29A와 AtNCED3 프로모터를 포함하는 스트레스 유도성 발현 binary 벡터들은 고염, ABA, MV 또는 저온과 같은 비생물성 스트레스에 의한 유전자의 이소성 발현에 유용하였다. 본 연구에서 제작된 binary 벡터들은 표적 유전자의 이소성 발현을 통해 유전자의 생물학적 기능연구, 분자생물학적작용 기작 연구에 유용하게 사용될 것으로 사료된다.

Keywords

References

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