Construction of specific erythromycin resistance mutations in the temperate lactococcal bacteriophage TP901-1 and their use in studies of phage biology

Koch, Birgit; Christiansen, Bettina; Evison, Tim; Vogensen, Finn K.; Hammer, Karin

doi:10.1128/aem.63.6.2439-2441.1997

Cited by 15 publications

(10 citation statements)

References 17 publications

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“…In a more sensitive infection assay, lysogenization frequencies of a variant of phage TP901-1, i.e. TP901-BC1034, were measured (Koch et al, 1997). The TP901-BC1034 phage contains an erythromycin resistance marker within the orf55 gene, which has no impact on its ability to perform either lytic growth or lysogenization on L. lactis subsp.…”

Section: Resultsmentioning

confidence: 99%

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Temperate phages TP901-1 and ÏLC3, belonging to the P335 species, apparently use different pathways for DNA injection inLactococcus lactissubsp.cremoris3107

Breum

Neve²,

Heller³

et al. 2007

FEMS Microbiology Letters

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Five mutants of Lactococcus lactis subsp. cremoris 3107 resistant to phage TP901-1 were obtained after treatment with ethyl methanesulfonate. Two of the mutants were also resistant to phage phiLC3. The remaining three mutants were as sensitive as 3107. Mutants E46 and E100 did not adsorb the two phages. Mutants E119, E121 and E126 adsorbed phage phiLC3 as well as 3107 but phage TP901-1 with significantly reduced efficiency. All, except E46, could be lysogenized with phage TP901-BC1034, a derivative of TP901-1 harboring an erythromycin-resistance marker. However, the lysogenization frequency was 10(3)-10(4) fold higher for 3107 than for the mutants. Mitomycin C induction of lysogenized mutants 3107 indicated that phage propagation was not affected in these four mutants. Electron microscopy and analysis of total DNA of infected cells showed that DNA was liberated from the phage particle during infection of strain 3107 with TP901-1 and that intracellular phage DNA replication occurred. This was not the case for mutants E121 and E126. This strongly suggests that some step starting with triggering DNA release and ending with DNA injection is impaired during infection with TP901-1. As such impairment was not seen when infecting E119, E121 and E126 with phiLC3, we conclude that TP901-1 and phiLC3 either are differently triggered by their receptor or utilize different pathways of injection.

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Section: Resultsmentioning

confidence: 99%

“…The EMS mutagenized L. lactis subsp. cremoris 3107 strains were infected with phage TP901-BK2, which contains an erythromycin resistance (Em r ) marker and lysogenizes at a low frequency compared with the wild-type phage (Koch et al, 1997). EMS-treated L. lactis subsp.…”

Section: Bacteriophage Propagation and Assaysmentioning

confidence: 99%

Temperate phages TP901-1 and ÏLC3, belonging to the P335 species, apparently use different pathways for DNA injection inLactococcus lactissubsp.cremoris3107

Breum

Neve²,

Heller³

et al. 2007

FEMS Microbiology Letters

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“…Several improvements were made to optimize the host background for analysing mutations made in TP901-1 erm , a derivative of TP901-1 that is marked with an erythromycin-resistance (Em r ) cassette[ 76 ], which had been used to select a TP901-1 erm lysogenized derivative of strain L . lactis NZ9000 [ 13 ].…”

Section: Resultsmentioning

confidence: 99%

“…The phage mix was then tested for its ability to infect L . lactis 3107 by assessing the frequency of lysogeny, which was determined through acquisition of the adenine methylase-encoding gene present on the TP901-1 erm phage genome (or its mutant derivatives), conferring resistance to 5 μg/ml erythromycin [ 11 , 76 ]. This was performed by adding 50 μl of the phage mix to 450 μl of L .…”

Section: Methodsmentioning

confidence: 99%

Structure and Assembly of TP901-1 Virion Unveiled by Mutagenesis

et al. 2015

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Bacteriophages of the Siphoviridae family represent the most abundant viral morphology in the biosphere, yet many molecular aspects of their virion structure, assembly and associated functions remain to be unveiled. In this study, we present a comprehensive mutational and molecular analysis of the temperate Lactococcus lactis-infecting phage TP901-1. Fourteen mutations located within the structural module of TP901-1 were created; twelve mutations were designed to prevent full length translation of putative proteins by non-sense mutations, while two additional mutations caused aberrant protein production. Electron microscopy and Western blot analysis of mutant virion preparations, as well as in vitro assembly of phage mutant combinations, revealed the essential nature of many of the corresponding gene products and provided information on their biological function(s). Based on the information obtained, we propose a functional and assembly model of the TP901-1 Siphoviridae virion.

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“…Lactococcal bacteriophage TP901-1[ 13 ] and TP901-BC1034[ 14 ] were used. The TP901-BC1034 is a TP901-1 derivative, which encodes erythromycin resistance and was used to aid measurement of lysogenization.…”

Section: Methodsmentioning

confidence: 99%

Clear Plaque Mutants of Lactococcal Phage TP901-1

et al. 2016

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We report a method for obtaining turbid plaques of the lactococcal bacteriophage TP901-1 and its derivative TP901-BC1034. We have further used the method to isolate clear plaque mutants of this phage. Analysis of 8 such mutants that were unable to lysogenize the host included whole genome resequencing. Four of the mutants had different mutations in structural genes with no relation to the genetic switch. However all 8 mutants had a mutation in the cI repressor gene region. Three of these were located in the promoter and Shine-Dalgarno sequences and five in the N-terminal part of the encoded CI protein involved in the DNA binding. The conclusion is that cI is the only gene involved in clear plaque formation i.e. the CI protein is the determining factor for the lysogenic pathway and its maintenance in the lactococcal phage TP901-1.

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Construction of specific erythromycin resistance mutations in the temperate lactococcal bacteriophage TP901-1 and their use in studies of phage biology

Cited by 15 publications

References 17 publications

Temperate phages TP901-1 and ÏLC3, belonging to the P335 species, apparently use different pathways for DNA injection inLactococcus lactissubsp.cremoris3107

Temperate phages TP901-1 and ÏLC3, belonging to the P335 species, apparently use different pathways for DNA injection inLactococcus lactissubsp.cremoris3107

Structure and Assembly of TP901-1 Virion Unveiled by Mutagenesis

Clear Plaque Mutants of Lactococcal Phage TP901-1

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Construction of specific erythromycin resistance mutations in the temperate lactococcal bacteriophage TP901-1 and their use in studies of phage biology

Cited by 15 publications

References 17 publications

Temperate phages TP901-1 and ÏLC3, belonging to the P335 species, apparently use different pathways for DNA injection inLactococcus lactissubsp.cremoris3107

Temperate phages TP901-1 and ÏLC3, belonging to the P335 species, apparently use different pathways for DNA injection inLactococcus lactissubsp.cremoris3107

Structure and Assembly of TP901-1 Virion Unveiled by Mutagenesis

Clear Plaque Mutants of Lactococcal Phage TP901-1

Contact Info

Product

Resources

About

Temperate phages TP901-1 and ÏLC3, belonging to the P335 species, apparently use different pathways for DNA injection inLactococcus lactissubsp.cremoris3107

Temperate phages TP901-1 and ÏLC3, belonging to the P335 species, apparently use different pathways for DNA injection inLactococcus lactissubsp.cremoris3107