Taxonomic Differentiation of Bacteriophages of Lactococcus lactis by Electron Microscopy, DNA-DNA Hybridization, and Protein Profiles

Braun, Volkmar; Hertwig, Stefan; Neve, Horst; Geis, Arnold; Teuber, Michael

doi:10.1099/00221287-135-9-2551

Cited by 83 publications

(100 citation statements)

References 23 publications

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“…Comparison of the 4vML3 restriction map ( Fig. 1) with those obtained for other prolate-headed phages (Powell & Davidson, 1985;Braun et al, 1989;Powell et al, 1989) shows no striking similarities. Comparison of prolate-headed phages has previously shown no correlation between homology based on DNA hybridization and restriction maps (Powell et al, 1989).…”

Section: Discussionmentioning

confidence: 97%

Characterization of the prolate-headed lactococcal bacteriophage OvML3: location of the lysin gene and its DNA homology with other prolate-headed phages

Shearman

Hertwig

Teuber

et al. 1991

Journal of General Microbiology

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4vML3 is a virulent prolate-headed bacteriophage that attacks a genetically well-studied strain family including Lactococcus Iactis subsp. Iactis strains 712, C2 and ML3. A restriction map was constructed for &ML3 using a wide variety of restriction endonucleases. The DNA was highly refractory to in uitro restriction; this is a common feature of lytic lactococcal phages. Genome size was estimated as 23 kb, which is similar to the sizes of other phages of the same morphological group. The presence of heat-dissociable DNA fragments was noted in gel electrophoresis of restriction enzyme digests. Cohesive ends were confirmed by pretreating the DNA with T4 ligase, which rendered the composite fragment insensitive to heat. The phage &ML3 genome thus consists of linear and non-permuted double-stranded DNA with complementary cohesive ends. The lysin gene from this phage acts as a specific lysis agent against the lactic streptococci. This gene has been cloned and sequenced previously. Further specific subcloning of EcoRV fragments of &ML3 DNA has located the lysin gene in the central region of the genome, orientated from right to left. This location was confirmed by hybridization of a lysin gene probe to @ML3 DNA. The lysin gene probe showed homology to a number of other prolate-headed phages including Pool. The lysin gene of PO01 was shown to be located in the central region of its genome. However, the isometric phage P107 did not hybridize, in spite of encoding its own lysin gene.

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Section: Discussionmentioning

confidence: 97%

Characterization of the prolate-headed lactococcal bacteriophage OvML3: location of the lysin gene and its DNA homology with other prolate-headed phages

Shearman

Hertwig

Teuber

et al. 1991

Journal of General Microbiology

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“…The temperate bacteriophage TP901-1 was induced from Lactococcus lactis subsp. cremoris 901-1 (Braun et al, 1989) by the use of UV light as previously described (Christiansen et al, 1994). L. lactis subsp.…”

Section: Methodsmentioning

confidence: 99%

“…More recently, DNA sequence analysis of entire lactococcal phage genomes has been performed and so far the published reports encompass three virulent phages, the prolateheaded related phages c2 and bIL67, and the small isometric-headed phage skl, and one temperate phage, rlt, also having a small isometric head (Lubbers et al, 1995;Schouler et al, 1994;Chandry et al, 1997;van Sinderen et al, 1996). TP901-1 is a temperate lactococcal phage and like r l t it belongs to the P335 class of small isometric phages (Jarvis et al, 1991;Braun et al, 1989). This class of phages is particularly interesting since it contains both virulent and temperate phages.…”

Section: Introductionmentioning

confidence: 99%

Temporal transcription of the lactococcal temperate phage TP901-1 and DNA sequence of the early promoter region

Madsen

Hammer

1998

Microbiology

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Transcriptional analysis by Northern blotting identified clusters of early, middle and late transcribed regions of the temperate lactococcal bacteriophage TP901-1 during one-step growth experiments. The latent period was found to be 65 min and the burst size 40210. The eight early transcripts, all mapping in a 13 kb region adjacent to the attachment site of TP901-1, were present at maximal levels 10 min after infection. The four middle transcripts, observed at maximal levels 30 min after infection, are all located within a 2 kb region at the distal end of the early transcripts. The late class of transcripts were detected 40 min after infection and the amounts of these transcripts increased with time. The late transcripts were localized to the 13 kb region adjacent to the 2 kb middle transcribed region. The sequence of almost 4 kb of the early region was determined, allowing a detailed transcriptional map for the early region of which in total 6.4 kb was sequenced. Sequence analysis of the early region revealed two closely positioned but divergently orientated promoters, P , and P, in accordance with the orientation of the ORFs and the transcriptional map. Nine ORFs were found, and similarities to a phage repressor, a single-stranded DNA-binding protein, a topoisomerase, a Cro-like protein and two other phage proteins of unknown function were detected. The gene arrangement in the early transcribed region of TP901-1 thus consists of two transcriptional units: one from P, containing four genes, of which at least two (the integrase gene and putative repressor) are needed for lysogeny, and the divergent and longer transcriptional unit from P, presumably encoding functions required for the l y t i c life cycle. ORFs with homology to proteins involved in DNA replication were identified on the latter transcriptional unit.

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“…The two sensitive strains belong to different subspecies: DB0410 is a strain of L. lactis subsp. lactis biovar diacetylactis, used as a starter culture for production of cheese and cultured buttermilk (Curic et al, 1999), while 901-1 belongs to the cremoris subspecies (Braun et al, 1989). The property to utilize orotate seems therefore to be strain-dependent rather than species-dependent.…”

Section: Discussionmentioning

confidence: 99%

The orotate transporter encoded by oroP from Lactococcus lactis is required for orotate utilization and has utility as a food-grade selectable marker

2007

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The orotate transporter encoded by oroP from Lactococcus lactis is required for orotate utilization and has utility as a food-grade selectable marker A new lactococcal plasmid, pDBORO, was isolated from the Lactococcus lactis subsp. lactis biovar diacetylactis strain DB0410. This plasmid is responsible for the sensitivity of DB0410 to the toxic pyrimidine analogue 5-fluoroorotate. The complete nucleotide sequence has been determined and amounts to 16 404 bp. Of 15 ORFs encountered, three were found to be insertion sequence (IS) elements, identified as two IS946 and one IS982. Two ORFs are incomplete due to the insertion of an IS element in their C-terminal region. Homologues for four ORFs were found in the IL1403 sequence: the copB gene, coding for a copperpotassium-transporting ATPase B, and the ysbA, ysbB and ysbC genes. The structural organization of the pDBORO replication region is highly similar to other theta-replicating plasmids in both the cis-(repA) and trans-acting (repB and orfX) sequences. By plasmid deletion analysis and molecular cloning, a single locus on pDBORO was found to confer sensitivity to 5-fluoroorotate. It was identified as ysbC, but renamed oroP in order to reflect its function. The oroP gene was found to be essential for the utilization of orotate as the sole pyrimidine source in a strain deficient in pyrimidine de novo synthesis. The amino acid sequence encoded by the ORF showed the characteristic features of a membrane protein. Therefore, oroP most probably encodes an orotate transporter. Surprisingly, homologues of oroP could be identified in the genomes of both L. lactis MG1363 and L. lactis IL1403 despite the fact that these strains were unable to significantly utilize orotate. Cloning of oroP in Escherichia coli and Bacillus subtilis showed that the orotate transport phenotype could be transformed to both organisms. The findings presented indicate that oroP can be used as a powerful, food-grade selection/ counterselection marker in many different organisms. INTRODUCTIONLactococcus lactis strains are widely used in the dairy industry as starter cultures in cheese fermentation. All L. lactis species isolated so far from starter cultures contain a number of different plasmids. These plasmids are thought to have been acquired during adaptation, and many plasmid-encoded functions have been related to or are essential for growth in milk. Examples are genes involved in lactose transport and metabolism , genes encoding proteinases necessary for casein degradation McKay et al., 1976) and di-and oligopeptide transport genes (Yu et al., 1995a(Yu et al., , 1996.Pyrimidine metabolism has been extensively studied in L. lactis in our laboratory (Andersen et al., 1996;Jorgensen et al., 2003Jorgensen et al., , 2004Kilstrup & Martinussen, 1998;Martinussen et al., , 2001Martinussen et al., , 2003Wadskov-Hansen et al., 2000, 2001, and a review of nucleotide metabolism in lactic acid bacteria has recently been published (Kilstrup et al., 2005). In L. lactis, pyrimidines can be synthesized de novo or ...

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Taxonomic Differentiation of Bacteriophages of Lactococcus lactis by Electron Microscopy, DNA-DNA Hybridization, and Protein Profiles

Cited by 83 publications

References 23 publications

Characterization of the prolate-headed lactococcal bacteriophage OvML3: location of the lysin gene and its DNA homology with other prolate-headed phages

Characterization of the prolate-headed lactococcal bacteriophage OvML3: location of the lysin gene and its DNA homology with other prolate-headed phages

Temporal transcription of the lactococcal temperate phage TP901-1 and DNA sequence of the early promoter region

The orotate transporter encoded by oroP from Lactococcus lactis is required for orotate utilization and has utility as a food-grade selectable marker

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