Abstract
To investigate the distribution of the hemolysin II determinant among strains of Bacillus cereus and Bacillus thuringiensis, thirteen strains of B. cereus and fourteen strains of B. thuringiensis strains were tested for hybridization of their chromosomal DNAs with a DNA probe containing the B. cereus hemolysin II gene. In addition, the production of hemolysin II, whose activity is not inhibited by cholesterol, was tested. The presence (absence) of the hybridization response in the microorganism's genome correlated with the presence (absence) of cholesterol-unaffected hemolysin production. Only four out of thirteen B. cereus strains were found to give a positive response in hybridization experiments, whereas thirteen out of fourteen B. thuringiensis strains responded positively. DNAs from ten B. thuringiensis strains contained a 3.5 kb EcoRV fragment, which hybridized with the B. cereus hemolysin II gene probe. The 3.5 kb EcoRV DNA fragment from one of these strains (B. thuringiensis VKM-B1555) was cloned and expressed in Escherichia coli cells. The hemolysin encoded by the cloned DNA fragment was not inhibited by cholesterol and possessed all other properties of B. cereus hemolysin II. The obtained data clearly show limited distribution of hemolysin II among B. cereus strains and demonstrate that hemolysin II is more characteristic of B. thuringiensis than B. cereus.
Similar content being viewed by others
References
Adler HI, Fisher WD, Cohen A, Hardigree AA (1967) Miniature Escherichia coli cells deficient in DNA. Proc Natl Acad Sci USA 57: 321–326.
Bernheimer AW (1988) Assay of hemolytic toxins. Methods Enzymol 165: 213–217.
Bernheimer A, Grushoff P (1967) Extracellular hemolysins of aerobic sporogenic Bacilli. J Bacteriol 93: 1541–1543.
Coolbaugh JC, Williams RP (1978) Production and characterization of two hemolysins of Bacillus cereus. Can J Microbiol 24: 1289–1295.
Cowell JL, Grushoff-Kosyk PS, Bernheimer AW (1976) Purification of cereolysin and the electrophoretic separation of the active (reduced) and inactive (oxidized) forms of the purified toxin. Infect Immun 14: 144–154.
Gavrilenko IV, Baida GE, Karpov AV, Kuzmin NP (1993) Nucleotide sequence for phospholipase C and sphingomyelinase genes from Bacillus cereus VKM-B164. Bioorg Chem 19: 133–138 [English translation of Bioorganicheskaya Khimia].
Gilmore MS, Cruz-Rodz AL, Leimeister-Wachter M, Kreft J, Goebel W (1989) A Bacillus cereus cytolytic determinant, cereolysin AB, which comprises the phospholipase C and sphingomyelinase genes: nucleotide sequence and genetic linkage. J Bacteriol 171: 744–753.
Honda T, Shiba A, Seo S, Yamamoto J, Matsuyama J, Miwatani T (1991) Identity of hemolysins produced by Bacillus thuringiensis and Bacillus cereus. FEMS Microbiol Lett 79: 205–210.
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277: 680–685.
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3: 208–218.
Pendleton IR, Bernheimer AW, Grushoff P (1973) Purification and partial characterization of hemolysins from Bacillus thuringiensis. J Invertebr Pathol 21: 131–135.
Sinev MA, Budarina ZhI, Gavrilenko IV, Tomashevski AYu, Kuzmin NP (1993) Evidence for the existence of Bacillus cereus hemolysin II: cloning of its genetic determinant. Mol Biol 27: 1218–1224 [English translation of Molekulyarnaya Biologiya].
Stoker NG, Pratt JM, Holland IB (1984) The minicell system. In: Hames BD, Higgins SJ (eds) Transcription and translation. A practical approach. IRL Press, Oxford, pp 164–171.
Tomita M, Taguchi IR, Ikezawa H (1982) Molecular properties and kinetic studies on sphingomyelinase of Bacillus cereus. Biochim Biophys Acta 704: 90–99.
Turnbull PCB (1986) Bacillus cereus toxins. In: Dorner F, Drews J (eds) Pharmacology of bacterial toxins. Pergamon Press, Oxford, pp 397–448.
Yamada A, Tsukagoshi N, Udaka S, Sasaki T, Makino S, Nakamura S, Little C, Tomita M, Ikezawa H (1988) Nucleotide sequence and expression in Escherichia coli of the gene coding for sphingomyelinase of Bacillus cereus. Eur J Biochem 175: 213–220.
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13 mp18 and pUC19 vectors. Gene 33: 103–119.
Zaitsev EN, Zaitseva EM, Bakhlanova IV, Gorelov VN, Kuzmin NP, Kryukov VM, Lanzov VA (1986) Cloning and characterization of recA gene from Pseudomonas aeruginosa. Genetics 22: 2721–2727 [English translation of Genetika].
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Budarina, Z.I., Sinev, M.A., Mayorov, S.G. et al. Hemolysin II is more characteristic of Bacillus thuringiensis than Bacillus cereus . Arch. Microbiol. 161, 252–257 (1994). https://doi.org/10.1007/BF00248701
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00248701