Bacillus thuringiensis 내에서 안정한 벡타를 이용한 cry1C 유전자의 발현

  • Choi, Soo-Keun (Bacterial Molecular Genetics Ru, Korea Research Institute of Bioscience and Biotechnology) ;
  • Oh, Keun-Hee (Bacterial Molecular Genetics Ru, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Jeong-Il (Bacterial Molecular Genetics Ru, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Seung-Hwan (Bacterial Molecular Genetics Ru, Korea Research Institute of Bioscience and Biotechnology)
  • 최수근 (생명공학연구소 미생물 분자유전학RU) ;
  • 오근희 (생명공학연구소 미생물 분자유전학RU) ;
  • 김정일 (생명공학연구소 미생물 분자유전학RU) ;
  • 박승환 (생명공학연구소 미생물 분자유전학RU)
  • Published : 1997.12.01

Abstract

During sporulation, Bacillus thuringiensis strains produce crystals consist of toxin proteins highly specific against insect pests. Their host specificities are desirable from a standpoint of environmental safety, but also limit market potential. Thus, development of improved Bacillus thuringiensis strains having broad host spectrum will contribute to increase its use. For the construction of Bacillus thuringiensis strain having broad host spectrum, we cloned cry1C gene encoding a toxin protein highly toxic against Spodoptera exigua from a B. thuringiensis isolate and constructed two recombinant plasmids, pUBClC and plC60. The plasmid PUBC1C has a replication origin of the natural plasmid pBC16 from B. cereus which is closely related species to B. thuringiensis, and the pBC16 was known to be replicated by rolling-circle mechanism. The plasmid pIC60 has a replication origin of a resident 60 MDa plasmid from B. thuringiensis subsp. kurstaki HD263, and it is believed that the pIC60 is replicated in a theta mode. The two plasmids were introduced into B. thuringiensis subsp. kurstaki cryB strain, and the transformed strains produced well-shaped bipyramidal crystals. We confirmed the expression of the cry1C gene by SDS-PAGE, and Western blotting. By investigating the segregational stability, it was found that the plasmid pIC60 is more stable than the pUBC1C.

Keywords

Bacillus thuringiensis;cryIC;Expression;Segregational stability

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