Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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DNA-mediated gene transfer in plant protoplasts

식물 원형질체에서의 marker gene 삽입

  • U, Zang-Kual (Applied Radioisotope Research Institute, Cheju National University) ;
  • Riu, Key-Zung (Applied Radioisotope Research Institute, Cheju National University) ;
  • So, In-Sup (Applied Radioisotope Research Institute, Cheju National University) ;
  • Hong, Kyung-Ae (Applied Radioisotope Research Institute, Cheju National University)
  • 유장걸 (제주대학교 방사능이용연구소) ;
  • 류기중 (제주대학교 방사능이용연구소) ;
  • 소인섭 (제주대학교 방사능이용연구소) ;
  • 홍경애 (제주대학교 방사능이용연구소)
  • Published : 1993.12.31
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Abstract

The neomycin phosphotransferase II gene (nptII) was introduced into geranium (Pelargonium zonale hybrids) protoplast by using PEG or electroporation method. The presence of the introduced DNA in the protoplast and the expressions of the gene in the transformed cells were examined. The presence of the nptII DNA in the protoplasts were detected by polymerase chain reaction. The expressions of nptII gene in the transformed cells were confirmed by the nptII assay.

Polyethylene glycol(PEG)법 또는 electroporation법으로 제라니움 원형질체에 neomycin phosphotransferase II(nptII) 유전자를 옮기고, 세포내에 도입된 nptII DNA의 존재유무와 발현여부를 조사하였다. Polymerase chain reaction(PCR)을 이용하여 검토한 결과, PEG법을 사용했을 때나 electroporation법을 사용했을 때 모두 세포내에 도입된 nptII DNA가 있음이 확인되었다. 또 이들 세포의 추출물을 전기영동하여 neomycin phosphotransferase 활성을 조사한 결과, 효소활성을 보이는 band가 검출되어 marker gene 으로 도입된 notII 유전자가 세포내에서 발현된다는 것이 확인되었다.

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