Taxonomic studies of Acinetobacter species based on outer membrane protein patterns.

Ino, Takeshi; Nishimura, Yûki

doi:10.2323/jgam.35.213

Cited by 9 publications

(5 citation statements)

References 26 publications

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“…DSM587) has been submitted to the GenBank database and assigned Accession Number X89709. In agreement with taxonomic studies based on outer membrane protein patterns (Ino and Nishimura, 1989), our determination of the 16S rDNA sequence of strain DSM587, A. calcoaceticus BD413-ivl10 (Juni and Janik, 1969), shows that it does not belong to the genospecies A. calcoaceticus as defined by Bouvet and Grimont (1986). We therefore refer to this strain as Acinetobacter sp.…”

Section: Nucleotide Sequence Accession Numbersupporting

confidence: 87%

System to study horizontal gene exchange among microorganisms without cultivation of recipients

Strätz

Mau

Timmis

1996

Molecular Microbiology

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Ribosomal RNA genes are characterized by highly conserved sequences and are present in multiple copies in most prokaryotic chromosomes. In principle, therefore, they might serve as sites for homologous recombination between unrelated microorganisms. Plasmids containing 23S ribosomal gene sequences, from different bacteria, which had been interrupted by insertion of a kanamycin-resistance gene, were used to transform Acinetobacter sp. DSM587 (former name: Acinetobacter calcoaceticus BD413-ivl10). In all cases, homologies between the 23S rRNA genes of phylogenetically distant bacteria and Acinetobacter sp. DSM587 were sufficient for replacement recombination events. The integration events, resulting in inactivation of any one of the seven rrn operons of Acinetobacter sp. DSM587, had no observable influence on cell growth. These results suggest the possibility of rRNA genes serving as natural vehicles for horizontal gene transfer. They also provide the basis of a novel strategy to analyse gene transfer without selection or cultivation of recipient cells. Because of the highly conserved structure of bacterial rrn operons, recombination events subsequent to gene transfer can be readily identified by polymerase chain reaction amplification of the recombinant sequence using a universal forward primer for the 16S rRNA gene and a reverse primer specific for the integrated marker gene.

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Section: Nucleotide Sequence Accession Numbersupporting

confidence: 87%

System to study horizontal gene exchange among microorganisms without cultivation of recipients

Strätz

Mau

Timmis

1996

Molecular Microbiology

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“…Classification of strains within the genus is complex and not completely resolved. According to some widely accepted classification schemes (Bouvet & Grimont, 1986 ;Bouvet et al, 1990;Ino & Nichimura, 1989), strain ADPl (or BD413) should not be designated A . calcoaceticus.…”

Section: U03772mentioning

confidence: 99%

IS 1236, a newly discovered member of the IS3 family, exhibits varied patterns of insertion into the Acinetobacter calcoaceticus chromosome

Gerischer

Da²,

Ln³

1996

Microbiology

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Analysis of spontaneous mutations in Acinetobacter calcoaceticus revealed a 1237 bp insertion sequence named IS7236 and possessing a nucleotide sequence resembling those of members of the IS3 family. The chromosome of A. calcoaceticus strain ADPl contains seven copies of IS7236 which appears to insert preferentially into pobR, the transcriptional activator of the structural gene for p-hydroxybenzoate hydroxylase. IS7236 creates tandem 3 bp DNA duplications flanking the sites of its insertion in pobR. Different duplication patterns are found following insertion of IS7236 into pcaH, a structural gene for protocatechuate 3,bdioxygenase. Therefore the insertion properties of IS7236 appear to be influenced by its DNA target. Amino acid sequences associated with the apparent transposase function have been conserved in ORFB of IS7236 whereas the presumed DNA-binding helix-turn-helix region of IS7236 ORFA exhibits substantial amino acid sequence divergence from its IS3 counterparts. IS7236 ORFA and ORFB coding sequences overlap considerably, and sequence evidence indicates mechanisms for ORFB expression in IS7236 may resemble those employed by other members of the IS3 family. Portions of the IS7236 terminal repeats exhibit substantial sequence divergence from other members of the IS3 family, but evolution appears to have conserved a mechanism preventing expression of the insertion sequence genes as a consequence of transcriptional readthrough.

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“…A number of recent studies have attempted to set up reliable identification schemes based on DNA homology, phenotypic characters, and comparison of cell envelope and outer membrane protein patterns, but such methods yield inconsistent results (1,10,11,12,23). The phenotypic characters of Acinetobacter species are susceptible to the conditions under which they are cultured (10).…”

mentioning

confidence: 99%

Sequencing of the rpoB Gene and Flanking Spacers for Molecular Identification of Acinetobacter Species

et al. 2006

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Acinetobacter species are defined on the basis of several phenotypic characters, results of DNA-DNA homology, and more recently, similarities or dissimilarities in 16S rRNA gene sequences. However, the 16S rRNA gene is not polymorphic enough to clearly distinguish all Acinetobacter species. We used an RNA polymerase ␤-subunit gene (rpoB)-based identification scheme for the delineation of species within the genus Acinetobacter, and towards that end, we determined the complete rpoB gene and flanking spacer (rplL-rpoB and rpoB-rpoC) sequences of the 17 reference strains of Acinetobacter species and 7 unnamed genomospecies. By using complete gene sequences (4,089 bp), we clearly separated all species and grouped them into different clusters. A phylogenetic tree constructed using these sequences was supported by bootstrap values higher than those obtained with 16S rRNA or the gyrB or recA gene. Four pairs of primers enabled us to amplify and sequence two highly polymorphic partial sequences (350 and 450 bp) of the rpoB gene. These and flanking spacers were designed and tested for rapid identification of the 17 reference strains of Acinetobacter species and 7 unnamed genomospecies. Each of these four variable sequences enabled us to delineate most species. Sequences of at least two polymorphic sequences should be used to distinguish Acinetobacter grimontii, Acinetobacter junii, Acinetobacter baylyi, and genomic species 9 from one another. Finally, 21 clinical isolates of Acinetobacter baumannii were tested for intraspecies relationships and assigned correctly to the same species by comparing the partial sequences of the rpoB gene and its flanking spacers.Acinetobacter species are ubiquitous in the environment and have emerged as important nosocomial pathogens (3). A propensity to tolerate drying (29, 30) and resistance to many commonly used antibiotics (28) are key factors in enabling the organism to survive and spread in the nosocomial environment. Furthermore, Acinetobacter species may serve as reservoirs of antibiotic-resistant genes, particularly in hospital environments (22).The genus Acinetobacter, originally proposed by Brisou and Prevot (6), comprises a collection of bacteria which show a great deal of phenotypic similarities. Currently, the genus Acinetobacter comprises many taxons based on hybridization groups and classified as genomovars and genomospecies. From an official taxonomic point of view, there are currently 17 Acinetobacter nomenspecies with standing in nomenclature. However, several other genomospecies have been proposed, mainly on the basis of DNA hybridization studies (7,20,21,28).Delineation of species within the genus Acinetobacter is often problematic (22). A number of recent studies have attempted to set up reliable identification schemes based on DNA homology, phenotypic characters, and comparison of cell envelope and outer membrane protein patterns, but such methods yield inconsistent results (1,10,11,12,23). The phenotypic characters of Acinetobacter species are susceptible to the condit...

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Taxonomic studies of Acinetobacter species based on outer membrane protein patterns.

Cited by 9 publications

References 26 publications

System to study horizontal gene exchange among microorganisms without cultivation of recipients

System to study horizontal gene exchange among microorganisms without cultivation of recipients

IS 1236, a newly discovered member of the IS3 family, exhibits varied patterns of insertion into the Acinetobacter calcoaceticus chromosome

Sequencing of the rpoB Gene and Flanking Spacers for Molecular Identification of Acinetobacter Species

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