Presence of plasmids in propionic acid bacteria

Panon, G

doi:10.1051/lait:198817

Cited by 6 publications

(5 citation statements)

References 13 publications

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“…Seventy-five Propionibacterium strains representing all four recognized species of dairy propionibacteria were screened for the presence of endogenous plasmids. In the majority of these strains no small endogenous plasmids could be found, in accordance with reports on other Propionibacterium strains (19,20,23). The following six strains were found to contain a 6-to 10-kb plasmid: P. acidipropionici ATCC 4875 (ϭ DSM 20272) and LMG 16447, P. jensenii LMG 16453, P. freudenreichii LMG 16545, P. freudenreichii subsp.…”

Section: Resultssupporting

confidence: 91%

Efficient Transformation System for Propionibacterium freudenreichii Based on a Novel Vector

Jore

Luijk

Luiten

et al. 2001

Appl Environ Microbiol

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A 3.6-kb endogenous plasmid was isolated from a Propionibacterium freudenreichii strain and sequenced completely. Based on homologies with plasmids from other bacteria, notably a plasmid from Mycobacterium, a region harboring putative replicative functions was defined. Outside this region two restriction enzyme recognition sites were used for insertion of an Escherichia coli-specific replicon and an erythromycin resistance gene for selection in Propionibacterium. Hybrid vectors obtained in this way replicated in both E. coli and P. freudenreichii. Whereas electroporation of P. freudenreichii with vector DNA isolated from an E. coli transformant yielded 10 to 30 colonies per g of DNA, use of vector DNA reisolated from a Propionibacterium transformant dramatically increased the efficiency of transformation (>10 8 colonies per g of DNA). It could be shown that restriction-modification was responsible for this effect. The high efficiency of the system described here permitted successful transformation of Propionibacterium with DNA ligation mixtures.The genus Propionibacterium can be divided into two groups, a group containing the classical (or dairy) propionibacteria and a group containing the cutaneous propionibacteria (6). Members of the first group, especially Propionibacterium freudenreichii, play an essential role in the manufacture of Swiss and related types of cheeses (12). Other industrial applications are found in the production of propionic acid and vitamin B 12 (5, 28). Of growing interest, but less well documented, are the probiotic properties ascribed to some propionibacterial strains (16,21).Strain improvement and, in general, study of this economically important group of bacteria would be greatly facilitated by the availability of a system for genetic modification. This report describes isolation and characterization of a 3.6-kb plasmid and successful use of this plasmid in the construction of a set of Escherichia coli-Propionibacterium shuttle vectors. Reproducible transformation of P. freudenreichii strains with these shuttle vectors was achieved by means of electroporation. MATERIALS AND METHODSBacterial strains and plasmids. Propionibacterium strains were obtained from the Belgian Coordinated Collections of Microorganisms/LMG (Ghent, Belgium), from the American Type Culture Collection (Rockville, Md.), and from the Deutsche Sammlung von Mikroorganismen (Braunschweig, Germany). P. freudenreichii subsp. freudenreichii VTB1 was obtained from DSM Food Specialties' industrial collection. For amplification of newly constructed shuttle vector DNA E. coli DH5␣ was used. pBluescript SKIIϩ was obtained from Stratagene (La Jolla, Calif.).Media and growth conditions. E. coli DH5␣ was cultivated at 37°C in L medium (24) supplemented with 50 g of ampicillin per ml if necessary. Propionibacteria were cultivated anaerobically at 30°C in MRS (7) or SLB medium (8) supplemented with an appropriate antibiotic when plasmid isolation was to be performed or in SLB medium when electroporation was to be performed.Isolatio...

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Section: Resultssupporting

confidence: 91%

Efficient Transformation System for Propionibacterium freudenreichii Based on a Novel Vector

Jore

Luijk

Luiten

et al. 2001

Appl Environ Microbiol

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“…Panon [32] screened 53 strains and found 20 strains harbouring each one or two plasmids (38%). It is noteworthy that a plasmid was isolated for the first time from a P. thoenii strain.…”

Section: Detection and Prevalence Of Propionibacterium Plasmidsmentioning

confidence: 99%

“…250 kb) isolated from a P. thoenii strain has been described so far [22]. Concerning P. thoenii strains only two more plasmids have been isolated from this species at all [32] (Stierli, unpublished).…”

Section: Detection and Prevalence Of Propionibacterium Plasmidsmentioning

confidence: 99%

“…The latter was hampered by the scarcity of relatively small plasmids in Propionibacterium, suitable for vector construction [16,32,34,36]. Indications that the G+C-content of DNA of the selection marker chosen is of importance, are supported by the observation that the use of a number of relatively A+T-rich marker genes, including erythromycin and chloramphenicol resistance genes, utilised in transformation systems of other bacteria, proved unsuccessful in Propionibacterium [16,17,24].…”

Section: Prerequisites For High Efficiency Transformation Of Propionimentioning

confidence: 99%

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Genetics and molecular biology of propionibacteria

Luijk¹,

Stierli

Schwenninger

et al. 2002

Lait

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-Research in the genetics and molecular biology of propionibacteria is currently making much progress. In order to develop efficient DNA transfer systems for the genus Propionibacterium, dairy and environmental propionibacteria were screened for the presence of suitable plasmids as a first step. Following nucleotide sequence analysis, potential replication functions were identified on several Propionibacterium plasmids such as pLME106/pRGO1, p545 and pLME108. Furthermore, ppnA, the gene encoding the propionicin SM1, was detected on pLME106. Three of these plasmids which had been fused with antibiotic resistance selection markers (ermE, cml, hygB) originating from bacteria with high G+C DNA content were recently successfully used as Escherichia coliPropionibacterium shuttle vectors. DNA restriction/modification systems observed in propionibacteria have to be taken into account since successful DNA transformation at high rates (up to 10 8 Propionibacterium transformants/µg of DNA) succeeds only with plasmid DNA originating from propionibacteria with the same restriction/modification system(s) as the strain to be transformed, and not from E. coli hosts. The basis for an integrating vector has been set up after identification of a potential attP site and an adjacent integrase gene from a Propionibacterium phage/prophage system. Finally, approximately 30 gene sequences with attributed coding functions from propionibacteria are available on databases.

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“…The size of plasmids varied between 3.2 and 47 kb; smaller plasmids were not observed. Panon [24] detected plasmids in 38 % of a total of 53 strains, which included al! the propionibacterial species.…”

Section: Plasmid Profilesmentioning

confidence: 99%

Propionibacteria flora in Swiss raw milk from lowlands and Alps

Fessler

Casey

Puhan

1999

Lait

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-The propionic acid bacterial tlora in Swiss raw milk was investigated. Four hundred and fifty-three strains from lowland milk and 21 strains from Alpine raw milk were c1assified to the species level by protein profile analysis and restriction analysis of the 23S ribosomal ribonucleic acid (rRNA). Plasmid profiles and random amplified polymorphie deoxyribonucleic acid (RAPD) was used to differentiate the strains. The tlora was found to be extremely diverse. Ali four dairy Propionibacterium species were found in lowland raw milk: 71 % were P.freudenreichii, 19 % P.jensenii, 8 % P. acidipropionici and 2 % P. thoenii. P. acidipropionici was not found in alpine milk but P. freudenreichii made up 55 %, P. jensenii 15 % and P. thoenii 30 % of the total. Among 278 P.freudenreichii strains, 219 (79 %) different strains were identified by RAPD to the strain level. For the other species strain diversity was even greater. Only 30 % of the strains analyzed carried plasmids. The Swiss chee se industry has a large reservoir of Propionibacterium strains in raw milk for future applications and developments. © InralElsevier, Paris propionibacterium / milk / electrophoresis / PCR Résumé -La flore propionique dans le lait cru suisse en provenance de la plaine et des alpages.La flore propionique dans le lait cru suisse a été analysée. L'analyse des profils protéiques et l'analyse de restriction de l'ARNr 235 ont été faites avec 453 souches bactériennes du lait en provenance de la plaine et 21 souches du lait d'alpages, en vue de les classer en espèces. Leur contenu en plasmides et la méthode de l'amplification génique au hasard de l'ADN ont été utilisés pour la diffé-renciation des souches. La tlore propionique dans le lait cru suisse s'est avérée être extrêmement variée. Les quatre espèces laitières de Propionibacterium ont été trouvées dans le lait cru de plaine; 71 % étaient des P. f reudenreichii, 19 % des P. jensenii, 8 % des P. acidipropionici et 2 % des P. thoenii. Dans le lait cru d'alpage P. acidipropionici n'a pas été trouvé, P. freudenreichii formait 55 %, P.jensenii 15 % et P. thoenii 30 % des bactéries propioniques. Parmi les 278 souches de P.freudenreichii isolées 219 (79 %) souches différentes ont été identifiées par RAPD. La diversité chez les autres

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Presence of plasmids in propionic acid bacteria

Cited by 6 publications

References 13 publications

Efficient Transformation System for Propionibacterium freudenreichii Based on a Novel Vector

Efficient Transformation System for Propionibacterium freudenreichii Based on a Novel Vector

Genetics and molecular biology of propionibacteria

Propionibacteria flora in Swiss raw milk from lowlands and Alps

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