Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Characteristics of Hemolysin in Mosquitocidal Bacillus thuringiensis strain 21-2

모기 살충성 Bacillus thuringiensis 21-2균주의 용혈성 내독소 단백질의 특성

  • Published : 2002.09.01
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Abstract

To describe characteristics of a hemolysin in mosquitocidal Bacillus thuringiensis subsp. gyangiensis strain 21-2, Escherichia coli HB101 was transformed with a gene encoding hemolysin in the strain 21-2. Transformant 47 con-tained 2.5 kb DNA was selected by ELISA, immunoblot and DNA electrophoresis. Transformant 47-5 was recon-structed after digestion of the 2.5 kb DNA with Hind m. Transformant 47-5 contained 1.8 kb DNA and expressed 23 kDa Protein which had mosquitocidal activity to Aedes aegypti. The 23 kDa Protein itself in vitro didn't show hemolytic activity on human erythrocytes, but the protein had the activity after proteinase K treatment.

모기 살충성 Bacillus thuringiensis subspangiensis 21-2균주의 용혈성 내독소 단백질의 특성을 검토하고자 21-2균주의 용혈성을 가진 유전자를 Escherichia coli HBIO쎄 형질전환시켰다. 이들 중에서 형질전환 균주 47은 독소 단백질을 생산하며 2.5 kb DNA을 함유한다는 것을 효소항체법, immunoblot및 DNA전기영동법으로 확인하였다. 또한, 형질전환 균주 47-5는 2.5 kb DNA를 다시 Hind ll견 절단하여 pUC118 연결시켜서 조제하였다 그 결과형질전환 균주 47-5은 1.Bkb DNA를 함유하며, 23 kD꺼 독소 단백질을 발현하고, 발현된 독소 단백질은 Aedes aegypti모기 유충에 독성을 나타내었다. 또한 23 kDa의 내독소 단백질 그 자체로는 사람의 적혈구를 용해하지 못하였으나, proteinase K로 처리한 후에는 적혈구에 대해 용혈성을 나타내었다.

Keywords

References

  1. J. Bacteriol. v.177 How does Bacillus thuringiensis produce so much insecticide crystal protein? Agaisse,H.;D.Lereclus https://doi.org/10.1128/jb.177.21.6027-6032.1995
  2. J. Invertbrate Pathol. v.70 Natural isolates of Bacillus thuringiensis: worldwide distribution, characterization, and cativity against insect pests Bernhard, K.;P.;Jarrett;M. Meadows;J. Butt;D. J. ellis;G. M. Roberts;S. Pauli;P. Rogers;H. D. Buerges https://doi.org/10.1006/jipa.1997.4669
  3. Can. J. Microbiol. v.43 Investigation of natural strains of Bacillus thuringiensis in different biotopes throughout the world Chaufaux,J.;M.N.Gilois;I.Jehanno;C.Buisson https://doi.org/10.1139/m97-047
  4. Appl. Environ. Microbiol. v.50 Micro-lipid-drop-let encapsulation of Bacillus thuringiensis subsp. israelensis delta-endotoxin for control of mosquito larvae Cheung, P. Y.;B. D. Hammock
  5. Kor. J. Appl. Microbiol. Biotechnol. v.18 Immunological characterization of mosquitocidal delta-endotoxins from Bacillus thuringiensis subsp. darmstadiensis 73E10-2 Cheung,T.Y.;K.H.Kim
  6. J. Gen. Microbiol. v.134 Comparative toxicity of Bacillus thuringiensis var. israelensis crystal proteins in vivo and in vitro Chilcott, C. N.;D. J. Ellar
  7. FEMS Microbiol. Lett. v.131 Contribution of the individual components of the $\delta$-endotoxin crystal to the mosquitocidal activity of Bacillus thuringiensis subsp. isralensis Crickmore,N.;E.J.Bone;J.A.William;D.J.Ellar
  8. Nucleic Acids Res. v.15 Bacillus thuringiensis var. morrisoni strain PG14: nucleotide sequence of a gene encoding a 27 kDa crystal protein Earp, D. J.;D. J. Ellar https://doi.org/10.1093/nar/15.8.3619
  9. Appl. Environ. Microbiol. v.63 Influence of single versus multiple toxins of Bacillus thuringiensis subsp. israelensis on development of resistance in the mosquito Culex quinquefasciatus(Diptera: Culicidae) Georghiou,G.P.;M.C.Wirth
  10. Appl. Envrion. Microbiol. v.67 The Bacillus thuringiensis cyt genes for hemolytic endotoxins constitute a gene family Guerchicoff, a.;A. Delecluse;C. P. Rubinstein https://doi.org/10.1128/AEM.67.3.1090-1096.2001
  11. J. Bacteriol. v.169 Transformation of vegitative cells of Bacillus thuringiensis by plasmid DNA Heierson,A.;L.Ritva;H.G.Boman https://doi.org/10.1128/jb.169.3.1147-1152.1987
  12. Microbiol. Rev. v.53 Insecicidal crystal proteins of Bacillus thuringiensis Hoffe, H.;H. R. Whiteley
  13. Curr. Microbiol. v.29 The 23-kilodalton CytB protein is solely responsible for mosquito larvicidal activity of Bacillus thuringiensis serovar kyushuensis Ishii,T.;M.Ohba https://doi.org/10.1007/BF01575754
  14. Appl. Environ. Microbiol. v.68 Characterization of Cyt2Bc toxin from Bacillus thuringiensis subsp. medellin Juarez-Perez,V.;A.Guerchicoff;C.Rubinstein;A.Delecluse https://doi.org/10.1128/AEM.68.3.1228-1231.2002
  15. Appl. Environ. Microbiol. v.61 Isolation and identification of novel toxins from a new mosquitocidal isolate from Malaysia, Bacillus thuringiensis subsp. jegathesan Kawalek, M. D.;S. Benjamin;H. L. Lee;S. S. Gill
  16. Kor. J. Appl. Microbiol. Biotechnol. v.25 Isolation and characaterization of Bacillus sp. PY123 producing water-soluble yellow pigment Kim, J. Y.;K. H. Kim
  17. J. Microbiol. Biotechnol. v.6 Cloning of a Hemolytic Mosquitocidal Delta-endotoxin Gene (cyt) of Bacillus thuringiensis 73E10-2 (serotype 10) into Bacillus subtilis and characterization of the cyt Gene Product Kim,K.H.;M.Ohba;B.W.Kim
  18. Kor. J. Appl. Microbiol. Biotechnol. v.27 Characterization of a mosquitocidal delta-endotoxin from Bacillus thuringiensis subsp. guiyangiensis strain 21-2(H serotype 43) Kim, W. J.;K. H. Kim
  19. Biochim. Biophys. Acta. v.924 Colloid-osmotic lysis is a general feature of the mechanism of action of Bacillus thuringiensis δ-endotoxins with different specificity Knowles, B. H.;D. J. Ellar https://doi.org/10.1016/0304-4165(87)90167-X
  20. FEBS v.244 A cytolytic $\delta$-endotoxin from Bacillus thuringiensis var. israelensis forms cation-selective channels in planar lipid bilayers Knowles,B.H.;M.R.Blatt;M.Tester;J.M.Horsnell;J.Carroll;G.Menestrina;D.J.Ellar https://doi.org/10.1016/0014-5793(89)80540-X
  21. Proc. Roy. Soc. Lond. Ser. B. v.248 A broad spectrum cytolytic toxin from Bacillus thringiensis var. Kyushuensis Knowles, B. H.;P. J. White;C. N. Nicholl;D. J. Ellar https://doi.org/10.1098/rspb.1992.0035
  22. Microbiology v.140 Biochemical characterization of Bacillus cytolytic δ-endotoxins Koni, P. A.;D. J. Ellar
  23. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli,U.K. https://doi.org/10.1038/227680a0
  24. J. Biol. Chem. v.193 Protein measurement with the Folin phenol reagent Lowry, O. H.;H. J. Rosebroughl;A. L. Farr;r. J. Randall
  25. Molecular cloning a laboratory manual(2nd ed.) Sambrook,J.;E.F.Fritsch;R.Maniatis
  26. J. Appl. Microbiol. v.84 Delta-endotoxin proteins associated with spherical parasporal inclusions of the four Lepidoptera- specific Bacillus thuringiensis strains Wasano,N.;K.H.Kim;M.Ohba https://doi.org/10.1046/j.1365-2672.1998.00371.x
  27. Appl. Environ. Microbiol. v.64 Variable cross-resistance to Cry 11B from Bacillus thuringiensis subsp. jegathesan in Culex quinquefasciatus (Diptera: Culicidae) resistant to single or multiple toxins of Bacillus thuringiensis subsp. israelensis Wirth,M.C.;A.Delecluse;B.A.Federici;W.E.Walton
  28. Appl. Environ. Microbiol. v.66 Cyt1A from Bacillus thuringiensis synergizes of Bacillus sphaericus against Aedes aegypti (Diptera: Culicidae) Wirth,M.C.;B.A.Federici;W.E.Walton https://doi.org/10.1128/AEM.66.3.1093-1097.2000
  29. Proc. Natl. Acad. Sci. USA v.94 CytA enables CryⅣ endotoxins of Bacillus thuringiensis to overcome high levels of CryⅣ resistance in the mosquito Culex quinquefasciatus Wirth, M.C.;G. P. Georghiou;B. A. Federici https://doi.org/10.1073/pnas.94.20.10536
  30. Mol. Microbiol. v.13 Synergism of mosquitocidal toxicity between CytA and CryIVD proteins using inclusions produced from cloned gene of Bacillus thuringiensis Wu,D.;J.Johnson;B.A.Federici https://doi.org/10.1111/j.1365-2958.1994.tb00488.x
  31. Appl. Environ. Microbiol. v.57 Characterization of mosquitocidal activity of Bacillus thuringiensis subsp. fukuokaensis crystal proteins Yu, Y. M.;M. Ohba;S. S. Gill