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Inhibition of a conjugation-like gene transfer process in Bacillus thuringiensis subsp. israelensis by the anti-s-layer protein antibody

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Abstract

Extraction of the S-layer protein by treatment with 6 m urea revealed a high-molecularweight protein in the extracts obtained from Bacillus thuringiensis subsp. israelensis (B.t.i) strain 4Q2. This protein band was found to be absent in partially cured (4Q2-72) and completely cured (c4Q2-72) strains. The antibody toward this S-layer protein was prepared and used to locate its antigenic protein on B.t.i cells by using indirect immunofluorescence. Immunodiffusion reactions and Western blot analysis confirmed the specificity of the anti-S-layer protein antibody. It was found that the antibody against 4Q2 S-layer protein, inhibited plasmid transfer via a conjugationlike process between, B.t.i. strains 4Q2-16 and c4Q2-72. That is, the frequency of transfer of plasmid pBC16 was reduced from 9.7×10-6 in the absence of the antibody to less than 1.0×10-8 in the presence of the antibody. The antibody was also found to reduce the frequency of pBC16 plasmid transfer via a conjugation-like process between B.t.i. strains A084-16-194 and c4Q2-72 from 2.2×10-5 in the absence of the antibody to 1.2×10-6 in the presence of the antibody.

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Literature Cited

  1. Andrews RE, Bulla LA (1982) Toxin of spore forming bacteria. In: Levinson HS, Sonenshein AL, Tipper DJ (eds) spore III. Washington, DC: American Society for Microbiology, p 57

    Google Scholar 

  2. Andrews RE, Faust RM, Wabiko H, Raymond KC (1987) The biotechnology of Bacillus thuringiensis. CRC Crit Rev Biotechnol 6:163–232

    Google Scholar 

  3. Aronśon AI, Beckman W, Dunn P (1986) Bacillus thuringiensis and related insect pathogens. Microbiol Rev 50:1–24

    Google Scholar 

  4. Battisti L, Green BD, Thorne CB (1985) Mating system for transfer of plasmids among B. anthracis, B. cereus and B. thuringiensis. J Bacteriol 162:543–550

    Google Scholar 

  5. Baumeister W, Hegeral R (1986) Can S-layers make bacterial connexons? FEMS Microbiol Lett 36:119–125

    Google Scholar 

  6. Baumeister W, Karrenberg F, Rachel R, Engel A, Heggeler B, Saxton WO (1982) The major cell envelope protein of Microccus radiodurans (R1): structure and chemical characterization. Eur J Biochem 125:535–544

    Google Scholar 

  7. Baumeister W, Barth M, Hegerl R, Guckenberger R (1986) Three-dimensional structure of the regular surface layer (HPI layer) of Deinococcus radiodurans. J Mol Biol 187:241–253

    Google Scholar 

  8. Bone EJ, Ellar DJ (1989) Transformation of Bacillus thuringiensis by electroporation. FEMS Microbiol Lett 58:171–178

    Google Scholar 

  9. Boonpucknavig S (1981) Immunofluorescent technique. Bangkok: Mahidol University, pp 39–40

    Google Scholar 

  10. Chapman J, Carlton BC (1985) Conjugal plasmid transfer in Bacillus thuringiensis. In: Helinski DR, Cohen SN, Clewell DB, Jackson DA, Hollaender A (eds) Plasmid in bacteria. New York: Plenum Press, pp 453–467

    Google Scholar 

  11. Dean DH (1989) Bacillus genetic stock center: strains and data, 4th ed. Columbus, Ohio: The Ohio State University

    Google Scholar 

  12. Evenberg D, Versluis R, Lugtenberg B (1985) Biochemical and immunological characterization of the cell surface of the fish pathogenic bacterium Aeromonas salmonicida. Biochim Biophys Acta 815:233–244

    Google Scholar 

  13. Fischer HM, Luthy P, Schweitzer S (1984) Introduction of plasmid pC194 into Bacillus thuringiensis by protoplast transformation and plasmid transfer. Arch Microbiol 139:213–217

    Google Scholar 

  14. Gonzalez JM, Brown BJ, Carlton BC (1982) Transfer of Bacillus thuringiensis plasmids coding for Delta-endotoxin among strains of B. thuringiensis and B. cereus Proc Natl Acad Sci USA 79:6951–6955

    Google Scholar 

  15. Hastie AT, Brinton CC (1990) Isolation, characterization, and in vitro assembly of the tetragonally arrayed layer of Bacillus sphaericus. J Bacteriol 138:999–1009

    Google Scholar 

  16. Howard L, Tipper DJ (1973) A polypeptide bacteriophage receptor: modified cell wall protein subunits in bacteriophageresistant mutants of Bacillus sphaericus strain P-1. J Bacteriol 113:1491–1504

    Google Scholar 

  17. Hudson L, Hay FC (1980) Practical immunology, 2nd ed. Oxford: Blackwell Scientific Publications, pp 124–128

    Google Scholar 

  18. Ishiguro EE, Key WW, Ainsworth T, Chamberlain JB, Austen RA (1981) Loss of virulence during culture of Aeromonas salmonicida at high temperature. J Bacteriol 148:333–340

    Google Scholar 

  19. Jones JB, Bowers S, Stadtman TC (1977) Methanococcus vanniellis ultrastructure and sensitivity to detergents and antibiotics. J Bacteriol 130:1357–1363

    Google Scholar 

  20. Kay WW, Phipps BM, Ishiguro EE, Trust TJ (1985) Porphyrin binding by the surface array virulence protein of Aeromonas salmonicida. J Bacteriol 164:1332–1336

    Google Scholar 

  21. Koval SF, Hynes SH (1991) Effect of paracrystalline protein surface layers in predation by Bdellovibrio bacteriovorus. J Bacteriol 173:2244–2249

    Google Scholar 

  22. Kupcu Z, Marz L, Messner P, Sleytr UB (1984) Evidence for the glycoprotein nature of the crystalline cell wall surface layer of Bacillus stearothermophilus strain NRS 2004/3a. FEBS Lett 173:185–190

    Google Scholar 

  23. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  24. Lereclus D, Arantes O, Chaufauxe J, Lecadet M-M (1989) Transformations and expression of cloned delta-endotoxin gene in Bacillus thuringiensis. FEMS Microbiol Lett 60:211–218

    Google Scholar 

  25. Lewis LO, Yousten AA, Murray RGE (1987) Characterization of the surface protein layers of the mosquito pathogenic strains of Bacillus sphaericus. J Bacteriol 169:72–79

    Google Scholar 

  26. Loprasert S, Pantuwatana S, Bhumiratana A (1986) Transfer of plasmids pBC16 and pC194 into Bacillus thuringiensis subsp. israelensis. J Invert Pathol 48:325–334

    Google Scholar 

  27. Luckevich M, Beveridge TJ (1990) Characterization of a dynamic S-layer on Bacillus thuringiensis. J Bacteriol 172:6656–6667

    Google Scholar 

  28. Mahillon J, Chungjatupornchai W, Decock J, Dierickx S, Michiels F, Peferoen M, Joos H (1987) Transformation of Bacillus thuringiensis by electroporation. FEMS Microbiol Lett 60:205–210

    Google Scholar 

  29. Martin PA, Lohr JR, Dean DH (1981) Transformation of Bacillus thuringiensis protoplasts by plasmid deoxyribonucleic acid. J Bacteriol 145:980–983

    Google Scholar 

  30. McCoy EC, Doyle D, Burda K, Corbeil LB, Winter AJ (1975) Superficial antigens of Campylobacter (Vibrio) fetus: characterization of an antiphagocytic component. Infect Immun 11:517–525

    Google Scholar 

  31. Mescher MF, Strominger JL (1976) Structure (shape-maintaining) role of the cell surface glycoprotein of Halobacterium salinarum. Proc Natl Acad Sci USA 73:2687–2691

    Google Scholar 

  32. Miteva VI, Shivarova NI, Grigorova RT (1981) Transformation of Bacillus thuringiensis protoplasts by plasmid DNA. FEMS Microbiol Lett 12:253–256

    Google Scholar 

  33. Ouchterlony O (1970) Handbook of immunodiffusion and immunoelectrophoresis. Ann Arbor, Mich: Ann Arbor Press

    Google Scholar 

  34. Phipps BM, Huber R, Baumeister W (1991) The cell envelope of the hyperthermophilic archaebacterium Pyrobaculum organotrophum consists of two regularly arrayed protein layers: three dimensional structure of the outer layer. Mol Microbiol 5:253–265

    Google Scholar 

  35. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press

    Google Scholar 

  36. Sleytr UB, Messner P (1983) Crystalline surface layers on bacteria. Annu Rev Microbiol 37:311–339

    Google Scholar 

  37. Sleytr UB, Sara M (1986) Ultrafiltration membranes with uniform pores from crystalline bacterial cell envelope layers. Appl Microbiol Biotechnol 25:83–90

    Google Scholar 

  38. Stoeckenius W, Rowen R (1967) A morphological study of Halobacterium halobium and its lysis in media of low salt concentration. J Cell Biol 34:365–393

    Google Scholar 

  39. Taylor KA, Deatherage JF, Amos LA (1982) Structure of the S-layer of Sulfolobus acidocaldarius. Nature 299:840–842

    Google Scholar 

  40. Trust TT, Kay WW, Ishiguro EE (1983) Cell surface hydrophobicity and macrophage association of Aeromonas salmonicida. Curr Microbiol 9:315–318

    Google Scholar 

  41. Tsukagoshi N, Yamada H, Tsuboi A, Udaka S, Katsura I (1982) Hexagonal surface array in a protein-secreting bacterium, Bacillus brevis 47. Biochim Biophys Acta 693:134–142

    Google Scholar 

  42. Wahlberg J, Tynkkynen S, Taylor N, Votila J, Kussinen K, Kussinen M, Haggblom M, Vilejanen J, Villstedt R, Nurmiaho-Lassila E-L, Lounatmaa K (1987) A hovel type of cell wall structure with two periodic layers (S-layer) of a Bacillus sp. strain KL1. FEMS Microbiol Lett 40:75–79

    Google Scholar 

  43. Weiss RL (1974) Subunits cell wall of Sulfolobus acidocaldarius. J Bacteriol 118:275–284

    Google Scholar 

  44. Wieland F, Dompert W, Bernhardt G, Sumper M (1980) Halobacterial glycoprotein saccharides contain covalently linked sulphate. FEBS Lett 120:110–114

    Google Scholar 

  45. Wiwat C, panbangred W, Bhumiratana A (1980) Transfer of plasmids and chromosomal genes amongst subspecies of Bacillus thuringiensis. J Ind Microbiol 6:19–27

    Google Scholar 

  46. Word NS, Yousten AA, Howard L (1983) Regularly structured and non-regularly structured surface-layers of Bacillus sphaericus. FEMS Microbiol Lett 17:277–282

    Google Scholar 

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Pharmacy, Mahidol University, 10400, Bangkok, Thailand

    Chanpen Wiwat

  2. Department of Biotechnology, Faculty of Science, Mahidol University, 10400, Bangkok, Thailand

    Watanalai Panbangred, Skorn Mongkolsuk & Amaret Bhumiratana

  3. Department of Microbiology, Faculty of Science, Mahidol University, 10400, Bangkok, Thailand

    Somsak Pantuwatana

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  1. Chanpen Wiwat
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  2. Watanalai Panbangred
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  4. Somsak Pantuwatana
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  5. Amaret Bhumiratana
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Wiwat, C., Panbangred, W., Mongkolsuk, S. et al. Inhibition of a conjugation-like gene transfer process in Bacillus thuringiensis subsp. israelensis by the anti-s-layer protein antibody. Current Microbiology 30, 69–75 (1995). https://doi.org/10.1007/BF00294185

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