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

  • 제21권3호
  • /
  • Pages.151-156
  • /
  • 2001
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  • 2287-5824(pISSN)
  • /
  • 2287-5832(eISSN)
한국초지조사료학회 (The Korean Society of Grassland and Forage Science)
권호 표지

BcHSP17.6 유전자 도입에 의한 알팔파의 형질전환

Transformation of Alfalfa by BcHSP17.6 Gene using Agrobacterium tumefaciens

  • Kim, K.Y. (National Livestock Research Institute) ;
  • Sung, B.R. (National Livestock Research Institute) ;
  • Rim, Y.W. (National Livestock Research Institute) ;
  • Choi, G.J. (National Livestock Research Institute) ;
  • Lim, Y.C. (National Livestock Research Institute) ;
  • Jang, Y.S. (National Livestock Research Institute) ;
  • Seo, S. (National Livestock Research Institute) ;
  • Yoon, S.H. (National Livestock Research Institute) ;
  • Park, G.J. (National Livestock Research Institute) ;
  • Jo, J. (College of Agriculture, Kyungpook National University)
  • 발행 : 2001.09.01
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초록

내열성 유전자인 BcHSP17.6를 갖도록 제작한 발현벡터 pBKH4를 Agrobacterium tumefaciens LBA 4404에 도입후, Agrobacterium과 알팔파 캘러스의 공배양을 통해 감염시킨 캘러스를 $100{\mu}g/m{\ell}$의 kanamycin과 $500{\mu}g/m{\ell}$의 cefotaxim을 첨가한 SH-kc배지에서 배양하며 형질전환된 캘러스를 선발하였다. 식물체 재분화는 SH- nk-c, SH-sp-c, SH-11b-c, SH-1BA 배지에서 약 4개월간 배양하여 재분화를 완성하였으며, 재분화된 알팔파의 genomic DNA를 분리한 후, PCR 분석 및 Southern blot 분석을 실시하여 알팔파의 형질전환을 확인하였다.

This study was conducted to obtain the transformed alfalfa (Medicago sativa L.) plants with thermotolerance gene (BcHSP17.6) using Agrobacterium tumefaciens LBA4404 and we confirmed the transformed gene from the regenerated alfalfa plants. The expression vector, pBKH4, harboring BcHSP17.6 gene was used for production of transgenic alfalfa plants. In a process for transformation, the callus of alfalfa was cocultivated with Agrobacterium tumefaciens and transformed calli were selected on kanamycin-containing SH-3-kc medium to regenerate into into the plant. The complete transgenic alfalfa plants were produced by cultivation for about 4 months on several regeneration media, SH-nk-c, SH-l lb-c, SH-sp-c, and SH-IBA. The transgenic alfalfa plants were analyzed by isolation of genomic DNA and PCR/Southem blot.

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