Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Gene Silencing Induced by Cytosine Methylation in Transgenic Tomato

형질전환 토마토에서 Cytosine Methylation에 의한 유전자발현 억제

  • Jung, Seo-Hee (Plant Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Min, Sung-Ran (Plant Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, Soo-Young (Plant Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Park, Ji-Young (Plant Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Davarpanah, S Javad (Plant Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Chung, Hwa-Jee (Plant Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Jeon, Jae-Heung (Plant Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Liu, Jang-Ryol (Plant Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Jeong, Won-Joong (Plant Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • 정서희 (한국생명공학연구원, 식물유전체연구센터) ;
  • 민성란 (한국생명공학연구원, 식물유전체연구센터) ;
  • 이수영 (한국생명공학연구원, 식물유전체연구센터) ;
  • 박지영 (한국생명공학연구원, 식물유전체연구센터) ;
  • ;
  • 정화지 (한국생명공학연구원, 식물유전체연구센터) ;
  • 전재홍 (한국생명공학연구원, 식물유전체연구센터) ;
  • 유장렬 (한국생명공학연구원, 식물유전체연구센터) ;
  • 정원중 (한국생명공학연구원, 식물유전체연구센터)
  • Published : 2007.12.31
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Abstract

Transgene expression was analyzed in tomato plants. Four lines of neomycin phosphotransferase II gene (NPTII) and the trehalose biosynthetic fusion gene (TPSP) transformed $T_0$ plants showed kanamycin resistance on selection medium. However, the analysis of phenotype (kanamycin resistance) and mRNA expression in $T_1$ plants indicated that the expression of the NPTII and TPSP transgenes was down-regulated to an undetectable level in two independent lines 1 and 11. Southern analysis demonstrated that the lines 1 and 11 had multicopies of the transgenes, whereas the typical transgenic lines 2 and 10 had 1 or 2 copies. DNA methylation analysis using methylation sensitive enzyme detected accumulated CpG DNA methylation on TPSP coding region and CaMV35S promoter region in the line 11, but not the typical transgenic line 2. These results suggest that multicopy transgene in plants is attributed to down-regulation of the transgene expression via transcriptional gene silencing.

토마토 형질전환체에서 NPTII 및 TPSP 유전자의 발현양상을 조사하였다. 4개 line 토마토 형질전환체가 선발배지에서 kanamycin에 저항성을 보이며 생산되었다. 그러나, 1번 line과 11번 line의 후대에서 NPTII 유전자의 발현이 억제되었으며, Kanamycin 저항성을 보이지 못했다. Southern 분석 결과, 1번 11번 line은 여러 copy의 외래 유전자를 가지고 있었으며, 2번, 10번 line은 1-2 copy의 외래유전자를 가지고 있었다. 형질전환 line 11번은 methylation sensitive 제한효소처리 후 DNA methylation 분석 결과, TPSP coding부위 및 도입유전자의 발현을 조절하는 CaMV35S 프로모터 주위에서 CpG methylation이 축적되었음이 확인되었다. 따라서 여러 copy의 외래유전자를 가진 토마토 형질전환 line 11번에서 NPTII 유전자 및 TPSP유전자의 발현이 억제된 것은 transcriptional gene silencing 기작에 의한 것으로 추정되었다.

Keywords

References

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