Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Expression of Pea Superoxide Dismutase Gene in Transgenic Cucumber (Cucumis sativus L.) Plants

형질전환 오이(Cucumis sativus L.) 식물체에서 완두 Superoxide Dismutase 유전자의 발현

  • 김재훈 (생명공학연구소 식물생화학 Research Unit.) ;
  • 오승용 (생명공학연구소 식물생화학 Research Unit.) ;
  • 이행순 (생명공학연구소 식물생화학 Research Unit.) ;
  • 조만현 (충남농촌진흥원 원예과) ;
  • 이은모 (충남농촌진흥원 원예과) ;
  • 우인식 (충남농촌진흥원 원예과) ;
  • 곽상수 (생명공학연구소 식물생화학 Research Unit.)
  • Published : 1998.05.01
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Abstract

To develop the fruits of cucumber (Cucumis sativus L.) producing high yields of superoxide dismutase (SOD), the MnSOD cDNA from pea (Pisum sativum) under the control of the cauliflower mosaic virus 35S promoter was introduced into cucumber using Agrobacterium tumefaciens (strain LBA 4404)-mediated transformation. The kanamycin-resistant shoots were selected on the selection medium containing MS basal salt, 1.0 mg/L zeatin, 0.1 mg/L IAA, 300 mg/L claforan, and 100 mg/L kanamycin. After 6 weeks of culture on the selection medium, the shoots were transferred to MS medium containing 0.2 mg/L NAA to induce roots. PCR analysis using the primers for neomycin phosphotransferase (NPTII) gene revealed that three plantlets were transformed. The fruits of one transgenic plant had approximately 3.2-fold higher SOD activity than those of non-transgenic plants. MnSOD isoenzyme band was strongly detected on native gel in fruits of transgenic plants.

Superoxide dismutase (SOD)를 많이 생산하는 신기능성 오이(Cucumis sativus L.) 과실을 개발하기 위하여, 발아 7일째의 자엽절편에 완두콩(Pisum sativum) 유래 MnSOD 유전자를 Agrobacterium tumefeciens LBA4404의 매개로 형질 전환시켰다. 1mg/L zeatin, 0.1mg/L IAA, 300mg/L claforan, 100mg/L kanamycin이 포함된 선발배지에서 부정아를 유도한 후, 1주일 간격으로 계대배양하면서 kanamycin 저항성 형질전환체를 선발하였다. 선발배지 배양 6주 후 kanamycin 저항성을 가진 부정아만을 절단하여 0.2mg/L NAA가 함유된 발근배지에 옮겨 뿌리를 유도한 후, 화분에서 순화시켰다. 소식물체의 잎에서 DNA를 분리하여 PCR 로 분석한 결과, 3개의 식물체에서 선발마커유전자인 NPTII 유전자가 존재함을 확인하여 SOD 유전자가 오이에 도입됨을 확인할 수 있었다. 이중 한 개체의 과실에서 SOD 함량이 무처리개체에 비해 약 3.2배 높았으며 native gel 전기영동에서 도입한 SOD isoenzyme 밴드가 강하게 검출되었다.

Keywords

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