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


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.


Bacillus thuringiensis;cryIC;Expression;Segregational stability


  1. Gene v.108 Construction of cloning vectors for Bacillus thuringiensis Arantes,O.;D.Lereclus
  2. Biochem. J. v.270 The construction of Bacillus thuringiensis strains expression novel entomocidal delta-endotoxin combinations Crickmore,N.;C.Nicholls;D.J.Earp;T.C.Hodgman;D.J.Ellar
  3. Gene v.96 High efficiency transformation of Escherichia coli with plasmids Inoue Hiroaki;Hiroshi Nojima;Hiroroto Okayama
  4. FEMS Microbiol. Lett. v.146 Discrimination between Bacillus cereus and Bacillus thuringiensis using specific DNA probes based on variable regions of 16S rRNA. te Giffel MC;R.R.Beumer;N.Klijn;A.Wagendo;F.M.Rombouts
  5. Appl. Environ. Microbiol. v.61 Enhanced production of insecticidal proteins in Bacilus thuringiensis strans carrying an additional crystal protein gene in their chromosome Kalman,S.;Kristine L. Kiehne;Nicole Cooper;Mitra Shahabi Reynoso;Takashi Yamamoto
  6. Gene v.20 Characterization and properties of a novel plasmid vector for Bacillus thuringiensis displaying compatibility with host plasmids Gamel,P.H.;Jean-Christophe Piot
  7. Appl. Environ. Microbiol. v.56 Novel cloning vectors for Bacillus thuringiensis Baum,J.A.;M.Dolores;M.Coyle;Pearce Gibert;Christine S. Jany;Cynthia Gawron-Burke
  8. Arch. Microbiol. v.139 Introduction of plasmid pC194 into Bacillus thuringiensis by protoplast transformation at plasmid transfer Fisher,H.M.;P.Luthy;S.Schweitzer
  9. Mol. Genet. v.210 Recombination between short repeated sequence is more frequent in plasmids than in the chromosome of Bacillus subtilis Janniere,L.;S.D.Ehrlish
  10. Mol. Genet. v.226 Plasmid deletion formation between short direct repeats in Bacillus subtilis is stimulated by single-stranded rolling-circle replication intermediates Bron,S.;S.Holsappeel;G.Venema;B.P.H.Peeters
  11. Appl. Environ. Microbiol. v.59 Cloning of a novel cry1C-type gene from a strain of Bacillus thuringiensis subsp. galleriae Kalman,S.;Kristine L. Kiehne;L.John;Takashi Yamamoto
  12. Plasmid v.25 Characterizationand sequence analysis of a small plasmid from Bacillus thuringiensis var. kurstaki strain HD1-DIPEL McDowell,D.G.;N.H.Mann
  13. Nucleic Acids Research v.18 Nucleotide sequence of the tetracycline resistance gene of pBC16 from Bacillus cereus Palva,A.;Gabriele Vidgren;Marjo simonen;Helena Rintala;Paivi Laamanen
  14. Appl. environ. Microbiol. v.59 The mode of replication is a major factor in segregational plasmid instability in Lactococcus lactis Kiewiet,R.;Jan Kok;Jos F.M.L. Seegers;Gerard Venema;Sierd Bron
  15. Molecular cloning: a laboratory manual Maniatis,T.;E.F.Fritsch;J.Sambrook
  16. Mol. Gen. Genet. v.218 Efficient transformation of Bacillus thuringiensis and B. cereus via electrophoration: transformation of acrystalliferous strains with a cloned delta-endotoxin gene Schurter,W.;M.Geiser;D.Mathe
  17. J. Bacteriol. v.173 Characterization and comparative sequence analysis of replication origins from three large Baillus thuringiensis plasmids Baum,J.A.;M.Pearce Gelbert
  18. An. Microbiol. v.150 Construction of a bifunctional genetically labelled plasmid for Bacillus thuringiensis subsp. israelensis Miteva,V.I.;R.T.Grigorova
  19. Gene v.87 Structurally stable Bacillus subtilis cloning vectors Janniere L.C. Bruand;S.D.Ehrlish