Abstract
In Escherichia coli, the fliC gene encodes flagellin, the protein responsible for eliciting the immunological reaction in H serotyping. Here, the presence of the flagellin fliC gene was studied in 86 Bacillus thuringiensis strains encompassing 67 H serotypes. Nineteen strains from four additional species in the B. cereus sensu lato group, B. cereus, B. anthracis, B. mycoides, and B. weihenstephanensis, were added for comparison purposes. The fliC genes were amplified, cloned and their nucleotide sequences determined and translated into amino acid sequences. A bootstrapped neighbor-joining tree was generated from the alignment of the translated amino acid sequences of the amplicons. Although most B. thuringiensis H serotypes had different flagellin amino acid sequences, some different B. thuringiensis serovars shared identical flagellin amino acid sequences. In addition, although serovars from the same H serotype were sometimes found clustered together, several serovars from the same H serotype carried flagellins with sufficiently different amino acid sequences as to be located on distant clusters. No correlations could be established between flagellin (FliC) protein sequence diversity among B. thuringiensis H serotypes and H serotype diversity. These suggest that the B. thuringiensis fliC gene does not code for the flagellin copy responsible for eliciting the immunological reaction in H serotyping. In a previous study, the authors have shown that the B. thuringiensis hag gene codes for the flagellin copy responsible for eliciting the immunological reaction in H serotyping. It is proposed that the B. thuringiensis fliC gene studied here be renamed and that the so-called hag gene studied before be renamed fliC, both in accordance with the E. coli nomenclature.
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References
Ankarloo J, Zhang M-Y, Lövgren A (1996) Regulatory sequences of two flagellin genes in Bacillus thuringiensis subsp. alesti. Microbiology 142:315–320
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
de Barjac H, Bonnefoi A (1962) Essai de classification biochimique et sérologique de 24 souches de Bacillus du type B. thuringiensis. Entomophaga 7:5–31
Entomogenous Organism Research Group (1976) Bacillus thuringiensis “140,” a new variety without flagellum. Acta Microbiol Sin 16:12–16
Glare TR, O’Callaghan M (2000) Bacillus thuringiensis: biology, ecology and safety. John Wiley and Sons, Toronto, Canada
Höfte H, Whiteley HR (1989) Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev 53:242–255
Iino T, Komeda Y, Kutsukake K, Macnab RM, Matsumura P, Parkinson JS, Simon MI, Yamaguchi S (1988) New unified nomenclature for the flagellar genes of Escherichia coli and Salmonella typhimurium. Microbiol Rev 52:533–535
Jung Y-C, Mizuki E, Akao T, Côté J-C (2007) A novel Bacillus thuringiensis strain expresses cytocidal activity against human cancer cells. J Appl Microbiol, in press, doi:10.1111/j.1365-2672.2006.03260.x
Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163
Lecadet M-M, Frachon E, Cosmao Dumanoir V, Ripouteau H, Hamon S, Laurent P, Thiéry I (1999) Updating the H-antigen classification of Bacillus thuringiensis. J Appl Microbiol 86:660–672
Lövgren A, Zhang M-Y, Engström A, Landén R (1993) Identification of two expressed flagellin genes in the insect pathogen Bacillus thuringiensis subsp alesti. J Gen Microbiol 139:21–30
Macnab RM (1992) Genetics and biogenesis of bacterial flagella. Annu Rev Genet 26:131–158
Ratiner YA (1983) Presence of two structural genes determining antigenically different phase-specific flagellins in some Escherichia coli strains. FEMS Microbiol Lett 19:37–41
Ratiner YA (1987) Different alleles of the flagellin gene hagB in Escherichia coli standard H test strains. FEMS Microbiol Lett 48:97–104
Ratiner YA (1998) New flagellin-specifying genes in some Escherichia coli strains. J Bacteriol 180:979–984
Saitou N, Nei M 1987 The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Stephen D, Jones C, Schofield JP (1990) A rapid method for isolating high quality plasmid DNA suitable for DNA sequencing. Nucleic Acids Res 18:7463–7464
Thompson JD, Higgins DG, Gibson TJ (1994) Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680
Xu D, Côté J-C (2003) Phylogenetic relationships between Bacillus species and related genera inferred from comparison of 3′ end 16S rDNA and 5′ end 16S-23S ITS nucleotide sequences. Int J Syst Evol Microbiol 53:695–704
Xu D, Côté J-C (2006) Sequence diversity of the Bacillus thuringiensis and B cereus sensu lato flagellin (H antigen) protein: comparaison with H serotype diversity. Appl Environ Microbiol 72:4653–4662
Acknowledgments
We thank Monika Ehling-Schulz from the Microbial Ecology Group, Technical University of Munich, Freising, Germany for providing us with B. weihenstephanensis DNA. We also thank Huguette de Barjac, Marguerite-M. Lecadet, Emmanuel Frachon and Jean-François Charles formerly from the International Entomopathogenic Bacillus Centre, Institut Pasteur, Paris, France, Daniel R. Zeigler from the Bacillus Genetic Stock Center, The Ohio State University, Columbus, OH, USA, and Zdena Pacova from the Czech Collection of Microorganisms, Masaryk University, Brno, Czech Republic, for providing us with bacterial strains.
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Soufiane, B., Xu, D. & Côté, JC. Flagellin (FliC) protein sequence diversity among Bacillus thuringiensis does not correlate with H serotype diversity. Antonie van Leeuwenhoek 92, 449–461 (2007). https://doi.org/10.1007/s10482-007-9173-3
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DOI: https://doi.org/10.1007/s10482-007-9173-3