Abstract
PacI and SpeI restriction maps were obtained for the two chromosomes of each of the six species of the genus Brucella: B. melitensis, B. abortus, B. suis, B. canis, B. ovis, and B. neotomae. Three complementary techniques were used: hybridization with the two replicons as probes, cross-hybridization of restriction fragments, and a new mapping method. For each type strain, a unique I-SceI site was introduced in each of the two replicons, and the location of SpeI sites was determined by linearization at the unique site, partial digestion, and end labeling of the fragments. The restriction and genetic maps of the six species were highly conserved. However, numerous small insertions or deletions, ranging from 1 to 34 kb, were observed by comparison with the map of the reference strain of the genus, B. melitensis 16M. A 21-kb Spel fragment specific to B. ovis was found in the small chromosome of this species. A 640-kb inversion was demonstrated in the B. abortus small chromosome. All of these data allowed the construction of a phylogenetic tree, which reflects the traditional phenetic classification of the genus.
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- Allardet-Servent A., Bourg G., Ramuz M., Pages M., Bellis M., Roizes G. DNA polymorphism in strains of the genus Brucella. J Bacteriol. 1988 Oct;170(10):4603–4607. doi: 10.1128/jb.170.10.4603-4607.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Canard B., Saint-Joanis B., Cole S. T. Genomic diversity and organization of virulence genes in the pathogenic anaerobe Clostridium perfringens. Mol Microbiol. 1992 Jun;6(11):1421–1429. doi: 10.1111/j.1365-2958.1992.tb00862.x. [DOI] [PubMed] [Google Scholar]
- Carlson C. R., Caugant D. A., Kolstø A. B. Genotypic Diversity among Bacillus cereus and Bacillus thuringiensis Strains. Appl Environ Microbiol. 1994 Jun;60(6):1719–1725. doi: 10.1128/aem.60.6.1719-1725.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Casjens S., Delange M., Ley H. L., 3rd, Rosa P., Huang W. M. Linear chromosomes of Lyme disease agent spirochetes: genetic diversity and conservation of gene order. J Bacteriol. 1995 May;177(10):2769–2780. doi: 10.1128/jb.177.10.2769-2780.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Colleaux L., Rougeulle C., Avner P., Dujon B. Rapid physical mapping of YAC inserts by random integration of I-Sce I sites. Hum Mol Genet. 1993 Mar;2(3):265–271. doi: 10.1093/hmg/2.3.265. [DOI] [PubMed] [Google Scholar]
- Daniels D. L. The complete AvrII restriction map of the Escherichia coli genome and comparisons of several laboratory strains. Nucleic Acids Res. 1990 May 11;18(9):2649–2651. doi: 10.1093/nar/18.9.2649. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Denoel P. A., Zygmunt M. S., Weynants V., Tibor A., Lichtfouse B., Briffeuil P., Limet J. N., Letesson J. J. Cloning and sequencing of the bacterioferritin gene of Brucella melitensis 16M strain. FEBS Lett. 1995 Mar 20;361(2-3):238–242. doi: 10.1016/0014-5793(95)00189-g. [DOI] [PubMed] [Google Scholar]
- Ficht T. A., Husseinen H. S., Derr J., Bearden S. W. Species-specific sequences at the omp2 locus of Brucella type strains. Int J Syst Bacteriol. 1996 Jan;46(1):329–331. doi: 10.1099/00207713-46-1-329. [DOI] [PubMed] [Google Scholar]
- Fitch W. M., Margoliash E. Construction of phylogenetic trees. Science. 1967 Jan 20;155(3760):279–284. doi: 10.1126/science.155.3760.279. [DOI] [PubMed] [Google Scholar]
- Fonstein M., Nikolskaya T., Haselkorn R. High-resolution alignment of a 1-megabase-long genome region of three strains of Rhodobacter capsulatus. J Bacteriol. 1995 May;177(9):2368–2372. doi: 10.1128/jb.177.9.2368-2372.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grothues D., Tümmler B. New approaches in genome analysis by pulsed-field gel electrophoresis: application to the analysis of Pseudomonas species. Mol Microbiol. 1991 Nov;5(11):2763–2776. doi: 10.1111/j.1365-2958.1991.tb01985.x. [DOI] [PubMed] [Google Scholar]
- Halling S. M., Bricker B. J. Characterization and occurrence of two repeated palindromic DNA elements of Brucella spp.: Bru-RS1 and Bru-RS2. Mol Microbiol. 1994 Nov;14(4):681–689. doi: 10.1111/j.1365-2958.1994.tb01306.x. [DOI] [PubMed] [Google Scholar]
- Halling S. M., Tatum F. M., Bricker B. J. Sequence and characterization of an insertion sequence, IS711, from Brucella ovis. Gene. 1993 Oct 29;133(1):123–127. doi: 10.1016/0378-1119(93)90236-v. [DOI] [PubMed] [Google Scholar]
- Hemmen F., Weynants V., Scarcez T., Letesson J. J., Saman E. Cloning and sequence analysis of a newly identified Brucella abortus gene and serological evaluation of the 17-kilodalton antigen that it encodes. Clin Diagn Lab Immunol. 1995 May;2(3):263–267. doi: 10.1128/cdli.2.3.263-267.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hill C. W., Harnish B. W. Inversions between ribosomal RNA genes of Escherichia coli. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7069–7072. doi: 10.1073/pnas.78.11.7069. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hiller B., Frey B., Schumann W. Tn 5Map, a transposon for the rapid mapping of restriction sites in plasmids. FEMS Microbiol Lett. 1994 Jan 15;115(2-3):151–155. doi: 10.1111/j.1574-6968.1994.tb06630.x. [DOI] [PubMed] [Google Scholar]
- Jiang Q., Hiratsuka K., Taylor D. E. Variability of gene order in different Helicobacter pylori strains contributes to genome diversity. Mol Microbiol. 1996 May;20(4):833–842. doi: 10.1111/j.1365-2958.1996.tb02521.x. [DOI] [PubMed] [Google Scholar]
- Jumas-Bilak E., Maugard C., Michaux-Charachon S., Allardet-Servent A., Perrin A., O'Callaghan D., Ramuz M. Study of the organization of the genomes of Escherichia coli, Brucella melitensis and Agrobacterium tumefaciens by insertion of a unique restriction site. Microbiology. 1995 Oct;141(Pt 10):2425–2432. doi: 10.1099/13500872-141-10-2425. [DOI] [PubMed] [Google Scholar]
- Komoda Y., Enomoto M., Tominaga A. Large inversion in Escherichia coli K-12 1485IN between inversely oriented IS3 elements near lac and cdd. Genetics. 1991 Nov;129(3):639–645. doi: 10.1093/genetics/129.3.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ladefoged S. A., Christiansen G. Physical and genetic mapping of the genomes of five Mycoplasma hominis strains by pulsed-field gel electrophoresis. J Bacteriol. 1992 Apr;174(7):2199–2207. doi: 10.1128/jb.174.7.2199-2207.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Le Bourgeois P., Lautier M., Mata M., Ritzenthaler P. Physical and genetic map of the chromosome of Lactococcus lactis subsp. lactis IL1403. J Bacteriol. 1992 Nov;174(21):6752–6762. doi: 10.1128/jb.174.21.6752-6762.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Le Bourgeois P., Lautier M., van den Berghe L., Gasson M. J., Ritzenthaler P. Physical and genetic map of the Lactococcus lactis subsp. cremoris MG1363 chromosome: comparison with that of Lactococcus lactis subsp. lactis IL 1403 reveals a large genome inversion. J Bacteriol. 1995 May;177(10):2840–2850. doi: 10.1128/jb.177.10.2840-2850.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leblond P., Redenbach M., Cullum J. Physical map of the Streptomyces lividans 66 genome and comparison with that of the related strain Streptomyces coelicolor A3(2). J Bacteriol. 1993 Jun;175(11):3422–3429. doi: 10.1128/jb.175.11.3422-3429.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu S. L., Hessel A., Sanderson K. E. The XbaI-BlnI-CeuI genomic cleavage map of Salmonella enteritidis shows an inversion relative to Salmonella typhimurium LT2. Mol Microbiol. 1993 Nov;10(3):655–664. doi: 10.1111/j.1365-2958.1993.tb00937.x. [DOI] [PubMed] [Google Scholar]
- Liu S. L., Sanderson K. E. Rearrangements in the genome of the bacterium Salmonella typhi. Proc Natl Acad Sci U S A. 1995 Feb 14;92(4):1018–1022. doi: 10.1073/pnas.92.4.1018. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu S. L., Sanderson K. E. The chromosome of Salmonella paratyphi A is inverted by recombination between rrnH and rrnG. J Bacteriol. 1995 Nov;177(22):6585–6592. doi: 10.1128/jb.177.22.6585-6592.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Margolin W., Bramhill D., Long S. R. The dnaA gene of Rhizobium meliloti lies within an unusual gene arrangement. J Bacteriol. 1995 May;177(10):2892–2900. doi: 10.1128/jb.177.10.2892-2900.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Michaux S., Paillisson J., Carles-Nurit M. J., Bourg G., Allardet-Servent A., Ramuz M. Presence of two independent chromosomes in the Brucella melitensis 16M genome. J Bacteriol. 1993 Feb;175(3):701–705. doi: 10.1128/jb.175.3.701-705.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Monteilhet C., Perrin A., Thierry A., Colleaux L., Dujon B. Purification and characterization of the in vitro activity of I-Sce I, a novel and highly specific endonuclease encoded by a group I intron. Nucleic Acids Res. 1990 Mar 25;18(6):1407–1413. doi: 10.1093/nar/18.6.1407. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ojaimi C., Davidson B. E., Saint Girons I., Old I. G. Conservation of gene arrangement and an unusual organization of rRNA genes in the linear chromosomes of the Lyme disease spirochaetes Borrelia burgdorferi, B. garinii and B. afzelii. Microbiology. 1994 Nov;140(Pt 11):2931–2940. doi: 10.1099/13500872-140-11-2931. [DOI] [PubMed] [Google Scholar]
- Ouahrani S., Michaux S., Sri Widada J., Bourg G., Tournebize R., Ramuz M., Liautard J. P. Identification and sequence analysis of IS6501, an insertion sequence in Brucella spp.: relationship between genomic structure and the number of IS6501 copies. J Gen Microbiol. 1993 Dec;139(12):3265–3273. doi: 10.1099/00221287-139-12-3265. [DOI] [PubMed] [Google Scholar]
- Perkins J. D., Heath J. D., Sharma B. R., Weinstock G. M. XbaI and BlnI genomic cleavage maps of Escherichia coli K-12 strain MG1655 and comparative analysis of other strains. J Mol Biol. 1993 Jul 20;232(2):419–445. doi: 10.1006/jmbi.1993.1401. [DOI] [PubMed] [Google Scholar]
- Sangari F. J., García-Lobo J. M., Agüero J. The Brucella abortus vaccine strain B19 carries a deletion in the erythritol catabolic genes. FEMS Microbiol Lett. 1994 Sep 1;121(3):337–342. doi: 10.1111/j.1574-6968.1994.tb07123.x. [DOI] [PubMed] [Google Scholar]
- Smith H. O., Birnstiel M. L. A simple method for DNA restriction site mapping. Nucleic Acids Res. 1976 Sep;3(9):2387–2398. doi: 10.1093/nar/3.9.2387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tatum F. M., Cheville N. F., Morfitt D. Cloning, characterization and construction of htrA and htrA-like mutants of Brucella abortus and their survival in BALB/c mice. Microb Pathog. 1994 Jul;17(1):23–36. doi: 10.1006/mpat.1994.1049. [DOI] [PubMed] [Google Scholar]
- Tatum F. M., Morfitt D. C., Halling S. M. Construction of a Brucella abortus RecA mutant and its survival in mice. Microb Pathog. 1993 Mar;14(3):177–185. doi: 10.1006/mpat.1993.1018. [DOI] [PubMed] [Google Scholar]
- Thierry A., Dujon B. Nested chromosomal fragmentation in yeast using the meganuclease I-Sce I: a new method for physical mapping of eukaryotic genomes. Nucleic Acids Res. 1992 Nov 11;20(21):5625–5631. doi: 10.1093/nar/20.21.5625. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tibor A., Weynants V., Denoel P., Lichtfouse B., De Bolle X., Saman E., Limet J. N., Letesson J. J. Molecular cloning, nucleotide sequence, and occurrence of a 16.5-kilodalton outer membrane protein of Brucella abortus with similarity to pal lipoproteins. Infect Immun. 1994 Sep;62(9):3633–3639. doi: 10.1128/iai.62.9.3633-3639.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zuerner R. L., Herrmann J. L., Saint Girons I. Comparison of genetic maps for two Leptospira interrogans serovars provides evidence for two chromosomes and intraspecies heterogeneity. J Bacteriol. 1993 Sep;175(17):5445–5451. doi: 10.1128/jb.175.17.5445-5451.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
