Genetic transformation of Pseudomonas aeruginosa with extracellular DNA.

Hara, T.; Aumayr, Andrea; Ueda, Seinosuke

doi:10.2323/jgam.27.109

Cited by 9 publications

(12 citation statements)

References 5 publications

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“…The presence of transforming DNA in cultures of Neisseria (2), pneumococci (12) and Bacillus (14) has been well documented. Hara et al (8) reported that the extracellular DNA produced by P. aeruginosa strain KYU-1 has a transforming histidine gene. Although the accumulation of the extracellular DNA by P. aeruginosa is thought to be due to the release of DNA with or without cell lysis, the exact mechanism of the release and the biological role of the extracellular DNA produced by P. aeruginosa have not been clarified.…”

Section: Discussionmentioning

confidence: 99%

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Transformation by Extracellular DNA Produced by Pseudomonas aeruginosa

Muto

Goto

1986

Microbiology and Immunology

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Most Pseudomonas aeruginosa strains are capable of producing extracellular DNA. Very closely linked chromosomal markers (leu+ and trp+) were cotransferred to P. aeruginosa PAO1819 (leu9001, trp9008) by the extracellular DNA produced by P. aeruginosa strains IFO3445 and PAO1 at a frequency of 10-7 to 10-8. Treatment of the extracellular DNA with DNase, heating at 95 C or sonication completely destroyed its transforming ability. The R plasmid in the extracellular DNA produced by P. aeruginosa IFO3445 (RP4) or PAO2142 (RLb679) could be transferred to Escherichia coli ML4901 or P. aeruginosa PA01819. The resultant transformants showed identical resistance patterns in the respective donors, and the sizes of the DNAs of RLb679 and RP4 isolated from the transformants were the same as those in the respective donors. These results demonstrate that the extracellular DNA contains both chromosomal DNA and plasmid DNA, and that it exhibits transforming ability. This implies that transformation by the extracellular DNA produced by P. aeruginosa may occur in nature and this seems to be of clinical importance in view of the spread of R plasmids among pathogens.Pseudomonas aeruginosa is well known as an opportunistic pathogen. In addition, the organism has the unique biological character of producing large amounts of DNA which accumulates extracellularly (7, 16). The occurrence of extracellular DNA has also been reported for Neisseria (2), Bacillus subtilis (15), Pseudomonas fluorescens (7), Vibrio parahaemolyticus (7), Staphylococcus aureus (3), Staphylococcus epidermidis (3), and Alcaligenes faecalis (3). Moreover, transformation by extracellular DNA has been demonstrated in Neisseria (2), Streptococcus (12), and B. subtilis (14).The mean size of the extracellular DNA produced by P. aeruginosa is estimated to be about 4 Mdal (16), hence it is possible that the extracellular DNA of P. aeruginosa and other bacteria has transforming ability. Hara et al (8) reported that a chromosomal marker (his+) in P. aeruginosa KYU-1 was transferred by intra-and extracellular DNA.Therefore, we investigated whether the extracellular DNA produced by P. aeruginosa can transfer chromosomal markers in P. aeruginosa by using two very closely linked selective markers. We also examined whether any plasmid DNA is contained in the extracellular DNA by searching for gene expression of the 621

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Section: Discussionmentioning

confidence: 99%

“…Hara et al (8) reported that a chromosomal marker (his+) in P. aeruginosa KYU-1 was transferred by intra-and extracellular DNA.…”

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confidence: 99%

Transformation by Extracellular DNA Produced by Pseudomonas aeruginosa

Muto

Goto

1986

Microbiology and Immunology

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“…In our previous papers (8)(9)(10)(11)(12), we reported that r-glutamyltranspeptidase (r-GTP) activity mainly participates in the biosynthesis of polyglutamate (PGA) in B. subtilis (natto) Asahikawa, isolated from the commercial product, "natto," and that the function of the 5.7-kilobases plasmid from the bacteria is in PGA production. Furthermore, we constructed the restriction endonuclease cleavage map of the plasmid (13).…”

Section: Vol 29mentioning

confidence: 99%

Identification of plasmids linked with polyglutamate production in Bacillus subtilis (natto).

Hara

Zhang

Ueda

1983

J. Gen. Appl. Microbiol.

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“…(I) Growth was determined by measuring the optical density (OD) at 660nm of appropriately diluted culture broth and was converted to dry weight as described in the previous paper. 1J (2) DNA samples of fibrous precipitate obtained by adding two volumes of 95% ethanol to the supernatant of culture broth were subjected to DNA assay by the meth09 of Dische 4J with calf thymus DNA as a standard. (3) RNA as a 75% ethanol-insoluble precipitate was determined by orcinol 5J with yeast RNA as standard.…”

Section: Microorganism the Microorganism Used In This Studymentioning

confidence: 99%

“…Moreover, DNA produced by this organism is active in transformation. 3 ) It has been known that various microorganisms excrete macromolecules such as protein and transforming DNA into the media, but less is known about the mechanism of DNA excretion. Extracellular DNA is found to originate from intracellular DNA and to be excreted under conditions in which lysis is not observed.…”

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confidence: 99%