Protein induced by bacteriophage T4 which is absent in Escherichia coli infected with nuclear disruption-deficient phage mutants

Koerner, James F.; Thies, Stephan; Snustad, D. Peter

doi:10.1128/jvi.31.2.506-513.1979

Cited by 6 publications

(3 citation statements)

References 33 publications

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“…Thus, a high level of Ndd provokes the rapid and complete disruption of the nucleoid, and no other T4 gene products are necessary. This correlates well with the fact that nucleoid disruption during wild-type T4 infection is achieved within a few minutes (18), when about 4,000 molecules of Ndd per cell are produced (11), and further suggests that Ndd be required to act at many sites simultaneously to bring about the destruction of the nucleoid. The previous observation that induction of Ndd expression produced only slow nuclear disruption is explained by the lower level of the protein in these experiments compared to that present after T4 infection (4).…”

supporting

confidence: 68%

“…The highly basic 17-kDa Ndd protein has no significant homology with any other known protein (5). About 4,000 Ndd molecules are produced by T4infected cells (11), but significantly lower levels of expression of the cloned ndd gene are nonetheless lethal to E. coli and induce a slow disorganization of the nucleoid (4). Ndd protein has little effect on bacterial gene expression but inhibits replication apparently by generating on the chromosome obstacles to progression of replication forks (4).…”

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confidence: 99%

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Ndd, the Bacteriophage T4 Protein That Disrupts the Escherichia coli Nucleoid, Has a DNA Binding Activity

1998

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Early in a bacteriophage T4 infection, the phage nddgene causes the rapid destruction of the structure of theEscherichia coli nucleoid. Even at very low levels, the Ndd protein is extremely toxic to cells. In uninfected E. coli, overexpression of the cloned ndd gene induces disruption of the nucleoid that is indistinguishable from that observed after T4 infection. A preliminary characterization of this protein indicates that it has a double-stranded DNA binding activity with a preference for bacterial DNA rather than phage T4 DNA. The targets of Ndd action may be the chromosomal sequences that determine the structure of the nucleoid.

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confidence: 68%

mentioning

confidence: 99%

Ndd, the Bacteriophage T4 Protein That Disrupts the Escherichia coli Nucleoid, Has a DNA Binding Activity

1998

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“…The actual number of molecules made each period was estimated by integrating the area under each curve and calculating ratios based on estimates in the literature. Koerner et al reported that T4 makes about 4000 molecules per cell of ndd, used to bind the host genome to the cell wall [ 20 ]. Burke et al estimated that 10,200 molecules/cell were made of gp45, involved in both DNA replication and late transcription [ 21 ].…”

Section: Figurementioning

confidence: 99%

From Host to Phage Metabolism: Hot Tales of Phage T4’s Takeover of E. coli

Kutter

Bryan

Ray

et al. 2018

Viruses

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The mechanisms by which bacteriophage T4 converts the metabolism of its E. coli host to one dedicated to progeny phage production was the subject of decades of intense research in many labs from the 1950s through the 1980s. Presently, a wide range of phages are starting to be used therapeutically and in many other applications, and also the range of phage sequence data available is skyrocketing. It is thus important to re-explore the extensive available data about the intricacies of the T4 infection process as summarized here, expand it to looking much more broadly at other genera of phages, and explore phage infections using newly-available modern techniques and a range of appropriate environmental conditions.

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The plasmid vectors, pBS2ndd and pBS3ndd, for versatile cloning with low background in Escherichia coli

Rotchanapreeda

Ngonsawan²,

Klomtun

et al. 2018

World J Microbiol Biotechnol

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For decades, diverse plasmid vectors have been continuously developed for molecular cloning of DNA fragment in the bacterial host cell Escherichia coli. Even with deliberate performances in vector preparation, the cloning approaches still face inevitable background colonies, or false positive clones, that may be arisen from intact or self-ligated plasmid molecules. To assist in such problem, two plasmids, pBS2ndd and pBS3ndd, which resistant to ampicillin and kanamycin respectively, were developed in this study as more advantageous cloning vector. The plasmids carry ndd, a lethal gene from bacteriophage T4 coding for nucleoid disruption protein that binds to the host chromosome and progressively kill the cell. The deadly toxicity of Ndd inhibits host cells that obtain intact or ndd-religated vector from growing, which results in low background and dramatically reduces the effort for selection of recombinants. Moreover, their identical multiple cloning site was designed to support various cloning strategies. Digestion of plasmids with XcmI allows for in vitro T/A ligation, while with EcoRV permits blunt-end ligation, with capability of blue-white colony screening. In vivo homologous recombination cloning is also utilizable by amplification of insert fragments using primers containing homology arms and transformation into capable E. coli strains. To demonstrate their advantages, the plasmids were used to clone PCR product samples for DNA sequencing with low-background and versatile cloning strategies. Such rapid and cost-effective cloning procedures are also proposed here. Finally, the cloning for protein expression with blue-white selection was also possible using egfp as a model regulated by lac and T7 promoters on the plasmid or other build-in promoters with the insert.

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Protein induced by bacteriophage T4 which is absent in Escherichia coli infected with nuclear disruption-deficient phage mutants

Cited by 6 publications

References 33 publications

Ndd, the Bacteriophage T4 Protein That Disrupts the Escherichia coli Nucleoid, Has a DNA Binding Activity

Ndd, the Bacteriophage T4 Protein That Disrupts the Escherichia coli Nucleoid, Has a DNA Binding Activity

From Host to Phage Metabolism: Hot Tales of Phage T4’s Takeover of E. coli

The plasmid vectors, pBS2ndd and pBS3ndd, for versatile cloning with low background in Escherichia coli

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