Isolation and characterization of two plasmids from
            <b>
              <i>Bifidobacterium longum</i>
            </b>

Park, M. S.; Lee, K. H.; Ji, Geun Eog

doi:10.1046/j.1472-765x.1997.00059.x

Cited by 32 publications

(19 citation statements)

References 3 publications

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“…Nevertheless, no more than two co-resident plasmids of a single strain have been fully characterized so far (Corneau et al 2004;Lee and O'Sullivan 2006;Park et al 1997). That is why we decided to perform a molecular characterization of the three plasmids harbored by B. longum NAL8, a strain isolated as a member of the predominant bifidobacterial population in a single fecal sample from a healthy elderly woman (Canzi et al 2005).…”

Section: Discussionmentioning

confidence: 99%

“…Shuttle vectors based on the ColE1 replication origin constitute the most commonly used system to isolate functional replicons of Bifidobacterium (Matteuzzi et al 1990;Rossi et al 1996;Matsumura et al 1997;Park et al 1997Park et al , 1999Tanaka et al 2005). However, when ColE1 was used successfully, it was obtained from pBR322, not from high-copy-number pUC vectors.…”

Section: Discussionmentioning

confidence: 99%

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Molecular characterization of Bifidobacterium longum biovar longum NAL8 plasmids and construction of a novel replicon screening system

Guglielmetti

Karp

Mora

et al. 2007

Appl Microbiol Biotechnol

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In this study, we performed molecular characterization and sequence analysis of three plasmids from the human intestinal isolate Bifidobacterium longum biovar longum NAL8 and developed a novel vector screening system. Plasmids pNAL8H (10 kb) and pNAL8M (4.9 kb) show close sequence similarity to and the same gene organization as the already characterized B. longum plasmids. The B. longum plasmid pNAC1 was identified as being most closely related to pNAL8L (3.5 kb). However, DNA sequence analysis suggested that direct repeat-rich sites could have promoted several recombination events to diversify the two plasmid molecules. We verified the likely rolling circle replication of plasmid pNAL8L and studied the phylogenetic relationship in all the Bifidobacterium plasmids fully sequenced to date based on in silico comparative sequence analysis of their replication proteins and iteron regions. Our transformation experiments confirmed that the ColE1 replication origin from high-copy-number pUC vectors could interfere with the replication apparatus of Bifidobacterium plasmids and give rise to false positive clones. As a result, we developed a system suitable for avoiding possible interference by other functional replication modules on the vector and for screening functional replicons from wildtype plasmids.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Molecular characterization of Bifidobacterium longum biovar longum NAL8 plasmids and construction of a novel replicon screening system

Guglielmetti

Karp

Mora

et al. 2007

Appl Microbiol Biotechnol

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“…Plasmids are not ubiquitous in bifidobacteria (332,392), and when present they are small, i.e., ranging from 1.5 kb to 15 kb. Completely sequenced plasmids from different B. longum biotype longum strains include pMB1 (378); pKJ36 and pKJ50 (332,333); pBLO1 (384); pNAC1, pNAC2, and pNAC3 (95); pDOJH10S and pDOJH10L (260); pTB6 (421); pB44 (GenBank accession number NC004443); and pNAL8 (162).…”

Section: Extrachromosomal Dna Elementsmentioning

confidence: 99%

“…Completely sequenced plasmids from different B. longum biotype longum strains include pMB1 (378); pKJ36 and pKJ50 (332,333); pBLO1 (384); pNAC1, pNAC2, and pNAC3 (95); pDOJH10S and pDOJH10L (260); pTB6 (421); pB44 (GenBank accession number NC004443); and pNAL8 (162). In addition, six plasmids from other bifidobacterial species have been sequenced: pVS809 from Bifidobacterium globosum (285), pCIBb1 from B. breve (327), pNBb1 from B. breve (GenBank accession number E17316), pAP1 from B. asteroides (GenBank accession number Y11549), pBC1 from Bifidobacterium catenulatum (5), and p4M from Bifidobacterium pseudocatenulatum (GenBank accession number NC003527).…”

Section: Extrachromosomal Dna Elementsmentioning

confidence: 99%

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Ventura

Canchaya

Tauch

et al. 2007

Microbiol Mol Biol Rev

936

638

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SUMMARY Actinobacteria constitute one of the largest phyla among Bacteria and represent gram-positive bacteria with a high G+C content in their DNA. This bacterial group includes microorganisms exhibiting a wide spectrum of morphologies, from coccoid to fragmenting hyphal forms, as well as possessing highly variable physiological and metabolic properties. Furthermore, Actinobacteria members have adopted different lifestyles, and can be pathogens (e.g., Corynebacterium, Mycobacterium, Nocardia, Tropheryma, and Propionibacterium), soil inhabitants (Streptomyces), plant commensals (Leifsonia), or gastrointestinal commensals (Bifidobacterium). The divergence of Actinobacteria from other bacteria is ancient, making it impossible to identify the phylogenetically closest bacterial group to Actinobacteria. Genome sequence analysis has revolutionized every aspect of bacterial biology by enhancing the understanding of the genetics, physiology, and evolutionary development of bacteria. Various actinobacterial genomes have been sequenced, revealing a wide genomic heterogeneity probably as a reflection of their biodiversity. This review provides an account of the recent explosion of actinobacterial genomics data and an attempt to place this in a biological and evolutionary context.

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“…Plasmid DNA from Bifidobacterium strains was prepared as described by Park et al [15]. Nucleotide sequences were determined using the BigDye terminator and the ABI377 system (PE Applied Biosystems, Foster City, CA, USA).…”

Section: General Cloning Techniques and Sequence Analysismentioning

confidence: 99%

Heterologous gene expression and secretion in Bifidobacterium longum

Park¹,

Seo²,

Kim

et al. 2005

Lait

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-Strains of the genus Bifidobacterium are used in fermented dairy products where they are supposed to exert health-promoting effects on the consumers. To study Bifidobacterium sp. properties, to improve the strains or to use them for safe delivery of vaccinal or anticarcinogenic polypeptides into the human digestive tract, it is strategically important to develop a system for heterologous expression and secretion. For this purpose, we constructed pBESAF2, a secretion vector based on the α-amylase expression and secretion signals (ESS) isolated from Bifidobacterium adolescentis. The signal peptide was analyzed in silico and its cleavage site was experimentally determined by Nterminal sequencing of the mature amylase. To validate this secretion vector, the phytase structural gene from Escherichia coli MC4100 was cloned downstream of the ESS and the resulting plasmid was introduced into Bifidobacterium longum MG1. Recombinant cells expressed phytase (with activity up to 21.1 U) and more than 95% of the phytase activity was found in the culture medium, suggesting that E. coli phytase can be produced and properly secreted and folded when expressed in B. longum. This is the first report of the development of a secretion vector for bifidobacteria and its use in a heterologous protein export in B. longum. This work is the first step towards the production of other proteins of food or therapeutic interest in bifidobacteria.Bifidobacterium / secretion / heterologous expression Résumé -Expression hétérologue et sécrétion chez Bifidobacterium longum. Certaines souches du genre Bifidobacterium sont utilisées dans des produits laitiers fermentés pour leurs potentiels effets bénéfiques sur la santé de l'hôte. Pour étudier les propriétés de Bifidobacterium sp., pour amé-liorer les souches ou pour les utiliser afin de délivrer des protéines vaccinales ou anticancéreuses dans le tube digestif de l'homme, il est stratégiquement primordial de développer un système d'expression et sécrétion hétérologue pour Bifidobacterium sp. À cette fin, nous avons construit pBESAF2, un vecteur de sécrétion utilisant des signaux d'expression et de sécrétion (SES) d'une amylase isolée de Bifidobacterium adolescentis. Le peptide signal a été analysé in silico et son site de clivage a été déterminé par séquençage amino-terminal de l'amylase mature. Pour valider ce vecteur de sécrétion, le gène spécifiant la phytase d'Escherichia coli MC1400 a été cloné en aval des SES et le plasmide résultant a été introduit dans B. longum MG1. Les bactéries recombinantes exprimaient la phytase (avec une activité allant jusqu'à 21,1 U) et plus de 95 % de l'activité phytase était détectée dans le surnageant de culture. Ce travail décrit la mise au point du premier vecteur de sécré-tion spécifiquement développé pour Bifidobacterium ainsi que son utilisation chez B. longum pour exprimer et sécréter une forme active de la phytase d'E. coli. Ce résultat ouvre la voie à la production d'autres protéines d'intérêt agro-alimentaire ou thérapeutique.Bifidobacterium / ...

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Isolation and characterization of two plasmids from Bifidobacterium longum

Cited by 32 publications

References 3 publications

Molecular characterization of Bifidobacterium longum biovar longum NAL8 plasmids and construction of a novel replicon screening system

Molecular characterization of Bifidobacterium longum biovar longum NAL8 plasmids and construction of a novel replicon screening system

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Heterologous gene expression and secretion in Bifidobacterium longum

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