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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Expression of recombinant protein from Oenanthe javanica DC. leaf tissues as a biofactory

Biofactory로서의 미나리 엽조직을 이용한 재조합단백질 발현

  • Shin, Dong-Il (Department of Biotechnology, Catholic University of Daegu) ;
  • Park, Hee-Sung (Department of Biotechnology, Catholic University of Daegu)
  • 신동일 (대구가톨릭대학교 생명공학과) ;
  • 박희성 (대구가톨릭대학교 생명공학과)
  • Published : 2008.12.31
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Abstract

Fresh Oenanthe javanica DC. leaves still attached to stem architecture were immersed in NaOH solution for 3 min before agroinfiltration and co-cultivation. MTT assay revealed that NaOH solution containing up to 0.7% was still safe for the leaf viability. Fluorometric GUS enzyme analysis showed that 0.5% NaOH-treated leaf tissues were efficiently transformed by vacuum infiltration for 20 min with Agrobacterium cells at a density of $OD_{600}=0.5$ to 1.0. These conditions worked well for the expression of HBsAg, which was confirmed by western blotting and ELISA.

미나리의 엽조직을 agroinfiltration 및 동시배양을 수행하기 전에 NaOH 용액에서 3 min 처리를 실시하였다. MTT를 이용한 세포사멸실험에서 0.7% NaOH 용액처리까지는 엽조직 세포활성에 안전한 것으로 판단되었다. GUS 효소활성의 형광분석 결과 0.5% NaOH를 처리한 미나리 엽조직에 대하여 Agrobacterium cells ($OD_{600}=0.5$에서 1.0)을 이용한 vacuum infiltration (20 min)을 실시할 경우 효율적 형질전환이 이루어짐을 알 수 있었다. 이러한 조건은 western blotting과 ELISA에 의한 HBsAg의 발현 검정에서 확인할 수 있었다.

Keywords

References

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