한국초지조사료학회지 (Journal of The Korean Society of Grassland and Forage Science)

  • 제26권2호
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  • Pages.83-90
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  • 2006
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  • 2287-5824(pISSN)
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  • 2287-5832(eISSN)
한국초지조사료학회 (The Korean Society of Grassland and Forage Science)
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E35S 프로모터 + AtNDPK2 유전자 도입에 의한 버즈풋 트레포일 (Lotus corniculatus L.) 형질전환체 생산

Production of Transgenic Birdsfoot Trefoil (Lotus corniculatus L.) Plants by Introduction of E35S Promoter + AtNDPK2 Gene

  • 발행 : 2006.06.01
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초록

환경스트레스에 내성을 갖는 버즈풋 트레포일(Lotus corniculatus L.) 형질전환체를 개발하기 위하여, AtNDPK 유전자가 E35S 프로모터에 의해 조절되도록 재조합한 발현벡터 pEN-K를 Agrobacterium 형질전환 방법으로 버즈풋 트레포일에 도입하였다. Agrobacterium과 공동배양한 버즈풋 트레포일 캘러스를 $100{\mu}g/m1$의 kanamycin 및 $500{\mu}g/m1$의 cefotaxim을 첨가한 SH-3-kc 배지에서 배양하며 식물체로 재분화시켰다. 재분화된 버즈풋 트레포일의 genomic DNA를 분리, PCR 및 Southern blot 분석을 실시한 결과, AtNDPK 유전자가 도입된 형질전환체의 경우 agarose gel 전기영동 및 X-ray 필름상에서 DNA band 및 hybridization signal을 확인할 수 있었으나, 형질전환되지 않은 대조구의 버즈풋 트레포일에서는 DNA band 및 hybridization signal이 관찰되지 않았다.

To develop transgenic birdsfoot trefoil (Lotus corniculatus L.) plants tolerant to environmental stress, Arabidopsis NDPK gene (AtNDPK) was introduced into birdsfoot trefoil plants using Agrobacterium-mediated transformation and expressed powerfully under the control of the E35S promoter. The expression vector, pEN-K was used for introduction of AtNDPK gene into birdsfoot trefoil plaits. The transformed calli were selected on kanamycin containing medium and then regenerated. The transformed birdsfoot trefoil plants were cultivated for 4 months on BOi2Y medium. Genomic DNA PCR and Southern blot analysis confirmed the incorporation of AtNDPK into the birdsfoot trefoil genome.

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참고문헌

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