Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Expression of Antibody Genes Specific for Human Hepatitis-B Virus in Transgenic Tabacco Plants

형질전환된 담배에서 사람 B형 간염바이러스 항체 유전자의 발현

  • Seok Yoon KWON (Genetic Engineering Research Institute, KIST) ;
  • Shin Je KIM (Genetic Engineering Research Institute, KIST Kumho Chemical Laboratories, Kprea Kumho Petrochemical Co., LTD.) ;
  • Hyo Jeong HONG (Genetic Engineering Research Institute, KIST) ;
  • Moon Hi HAN (Genetic Engineering Research Institute, KIST) ;
  • Chang Ho CHUNG (Kumho Chemical Laboratories, Kprea Kumho Petrochemical Co., LT) ;
  • Ho Sul LEE (Kumho Chemical Laboratories, Kprea Kumho Petrochemical Co., LTD) ;
  • Kyung Hee PAEK (Genetic Engineering Research Institute, KIST)
  • Published : 1994.11.01
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Abstract

Chimeric kappa chain and gamma chain cDNA clones (pCKS2 and pCHS2) of a monoclonal antibody specific for pre-S2 surface antigen of human hepatitis-B virus were ligated into Xbal site of plant expression vector pBKS-1. Plasmid DNA containing each of the chimeric gene were then mobilized from E, coli to Agrobacterium tumefaciens strain LBA4404. The chimeric antibody genes were then introduced into tobacco by Ti plasmid-mediated transformation. The putative Transformants were selected on medium containing kamaycin sulfate. Shoots that formed on shoot induction medium were analyzed by Western blot analysis for the expression of kappa-chain or gamma-chain genes. The Western blot analyses clearly showed that the introduced genes were stably expressed in transgenic plants.

사람 B형 간염 바이러스의 pre-S2 표면항원에 결합하는 키메라 항체 유전자(카파 및 감마사슬의 cDNA클론)를 식물체에서 발현시키기 위해 식물체 발현벡터인 pBKS-1에 XbaI 자리를 이용하여 클로닝하였다. 이들 유전자를 포함하는 대장균의 플라스미드 핵산을 추출하여 아그로박테리움에 형질전환 시켰다. 다음 담배의 조직절편과 아그로박테리움을 공동배양함으로써 담배의 형질 전환을 시도하였다. 카나마이신이 포함된 신초유기배지에서 나온 신초를 시료로 하여 Western blot을 실시함으로써 이들 유전자가 형질전환 담배에서 안정하게 발현됨을 확인하였다.

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