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Friday, July 31, 2020 | History

2 edition of Recombination of bacteriophage T7 DNA. found in the catalog.

Recombination of bacteriophage T7 DNA.

Donald Dah-Chen Lee

Recombination of bacteriophage T7 DNA.

by Donald Dah-Chen Lee

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  • 16 Currently reading

Published .
Written in English


The Physical Object
Pagination198 leaves.
Number of Pages198
ID Numbers
Open LibraryOL14731324M

Holliday junctions are resolved into recombinant duplex DNA species by a class of structure-specific endonucleases known as the Holliday junction-resolving enzymes. The primary cellular resolving enzyme in bacteria is RuvC, which is the main focus of this chapter. The author also talks about the RusA protein, which may act as an alternative to RuvC in some bacterial species, and attempts to Author: Malcolm F. White. In vitro repair of double-strand breaks accompanied by recombination in bacteriophage T7 DNA. By W Masker. Abstract. A double-strand break in a bacteriophage T7 genome significantly reduced the ability of that DNA to produce viable phage when the DNA was incubated in an in vitro DNA replication and packaging system. When a homologous piece of Cited by: 9.

Generalized transduction involving a bacterial virus (bacteriophage) and a donor bacterium. A second type of transduction is called specialized transduction. In this case, the lysogenic cycle ensues as before. When the phage DNA breaks away from the bacterial DNA, however, it may take with it a small amount of the bacterial DNA (perhaps 5 percent). Gene 6 protein of bacteriophage T7 has 5′-3′-exonuclease activity specific for duplex DNA. We have found that gene 6 protein also has flap endonuclease activity. The flap endonuclease activity is considerably weaker than the exonuclease activity.

Frazer J Rixon, Wah Chiu, in Advances in Protein Chemistry, 3 T4. The large DNA bacteriophages, typified by T4, have distinctive structures in which an icosahedrally organized head is connected at one vertex through a collar to a complex tail structure (Fig. 1E).Tails are found only in certain classes of bacteriophage and they seem to have evolved as an efficient tool for transferring. Bacteriophage, any of a group of viruses that infect bacteria. Bacteriophages were discovered independently by Frederick W. Twort in Great Britain () and Felix d’Herelle in France (). Thousands of varieties of phages exist. Certain types serve key roles in laboratory research.


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Recombination of bacteriophage T7 DNA by Donald Dah-Chen Lee Download PDF EPUB FB2

We have demonstrated recombination of bacteriophage T7 DNA in vitro. An extract of Escherichia coli B cells infected with wild-type T7(T7+) is incubated with mature DNA extracted from T7 phage. Packaging of the exogenous DNA within the phage head appears to be preceded by recombination of exogenously added DNA with DNA present in the by: ABSTRACT We have demonstrated recombination of bacteriophage T7 DNAin vitro.

An extract of Escherichia coli B cells infected with wild-type T7 (17+) is incubated withmatureDNAextracted ingofthe exogenousDNAwithinthephageheadappearsto bepreced-ed byrecombination of exogenously addedDNAwith DNA present in the by: We developed a simple, direct, physical assay to detect genetic recombination of bacteriophage T7 DNA in vitro.

In this assay two mature T7 DNA molecules, each having a unique restriction enzyme site, are incubated in the presence of a cell-free extract from T7-infected Escherichia coli by: 5. Most recombination following infection with T7 was found to coincide with the time of most rapid DNA synthesis, at about 20 min after infection at 30° in minimal medium.

Recombining DNA was investigated electron microscopically. Multiply branched DNA structures were observed after infection with T7 wild type, gene 3 −, gene 6 − and genes 3 −, 6 − phage, but not after infection with T7 Cited by:   We have demonstrated recombination of bacteriophage T7 DNA in vitro.

An extract of Escherichia coli B cells infected with wild-type T7(T7+) is incubated with mature DNA extracted from T7 phage. Packaging of the exogenous DNA within the phage head appears to be preceded by recombination of exogenously added DNA with DNA present in the by: A system capable of in vitro packaging of exogenous bacteriophage T7 DNA has been used to monitor the biological activity of DNA replicated in vitro.

This system has been used to follow the Recombination of bacteriophage T7 DNA. book of UV radiation on in vitro replication and by: 5.

VIROL () Genetic Recombination of Bacteriophage T7 DNA in Vitro II. Further Properties of the in Vitro Recombination-Packaging Reaction PAUL D. SADOWSKI Departments ofPathlology and Medical Genetics, University of Toronto, Toronto, Ontario M5S 1A8, Canada Accepted Decem Further properties of an in vitro recombination-packaging system for phage T7 DNA Cited by: The replication system of bacteriophage T7 is remarkable in that nucleotide genome is replicated over fold in a matter of minutes.

In order to accomplish this feat T7 has evolved an efficient and economical process for the replication of its DNA. The T7 replisome provides a model system to study DNA by: Genetic recombination in bacteriophage φX usually takes place early in the infection process and involves two parental replicative form (double-stranded) DNA molecules.

The host recA protein is required; none of the nine known φX cistron products is essential. The products of a single recombination event are nonreciprocal and by: nitiation of DNA replication in phage T7 chromosome.

The primary origin of phage T7 (position –) contains two T7 RNA polymerase promoters, ϕA and ϕB, an AT-rich region, and a. The experiments have implicated the products of gene 3 (T7 endonuclease), gene 4 (DNA replication protein), gene 5 (T7 DNA polymerase) and gene 6 (T7 exonuclease) in phage T7 genetic recombination.

During recent years a great deal of infor- mation has accumulated concerning DNA metabolism after phage T7 infection (7).Cited by: A recombination event, catalyzed by a phage coded enzyme, occurs between a particular site on the circularized phage DNA and a particular site on the host chromosome.

The result is the integration of the phage DNA into the host chromosome as illustrated in Figure 6. The initial step is the injection of T7 proteins from the phage particle to form a channel across the cell envelope that conducts DNA into the cell by a process that is apparently enzymatic Cited by: It turns out the early genes are transcribed by the bacterial host cell RNA polymerase.

Phage T7 late genes are transcribed by a phage-encoded RNA polymerase called GP1 (or T7 RNA Polymerase) that is totally unrelated to the bacterial host RNA polymerase.

The phage polymerase, "T7 polymerase", is the product of an early gene, called gene 1. Illegitimate recombination is a ubiquitous phenomenon and includes three types of events.

In the first class, rearrangements occur by recombination between short homologous sequences. A second class is associated with site-specific elements. A last class groups all rearrangements in which the newly linked sequences share less than 3 bp of homology and have no homology with known specific by: We have demonstrated recombination of bacteriophage T7 DNA in vitro.

An extract of Escherichia coli B cells infected with wild-type T7(T7+) is incubated with mature DNA extracted from T7 phage. Packaging of the exogenous DNA within the phage head appears to be preceded by recombination of exogenously added DNA with DNA present in the extracts.

A system capable of in vitro packaging of exogenous bacteriophage T7 DNA has been used to monitor the biological activity of DNA replicated in vitro. This system has been used to follow the effects of UV radiation on in vitro replication and recombination.

During the in vitro replication process, a considerable exchange of genetic information Author: W E Masker and N B Kuemmerle. When a homologous piece of T7 DNA (either a restriction fragment or T7 DNA cloned into a plasmid) that was by itself unable to form a complete phage was included in the reaction, the break was repaired to the extent that many more viable phage were by: 9.

A system capable of in vitro packaging of exogenous bacteriophage T7 DNA has been used to monitor the biological activity of DNA replicated in vitro.

This system has been used to follow the effects of UV radiation on in vitro replication and recombination. Mechanistic Studies of DNA Replication and Genetic Recombination emerged from a symposium on DNA replication and genetic recombination held from Marchin Keystone, Colorado.

The event featured 30 plenary session talks, 13 workshop discussion groups, and the poster sessions. RecBCD enzyme is centrally important in homologous recombination in Escherichia coli and is the source of ExoV activity.

Null alleles of either the recB or the recC genes, which encode the B and C subunits, respectively, manifest no recombination and none of the nuclease functions characteristic of the holoenzyme.

Loss of the D subunit, by a recD mutation, likewise results in loss of ExoV Cited by:   Lambda phage can be used for cloning of recombinant DNA, the use of its site specific recombinase (int) for the shuffling of cloned DNAs by the 'Gateway' method, and in recombineering.

The discovery and study of lambda phage led to an understanding of transduction which provides a major mechanism that can explain modes of evolution.Drexler, H.,Initiation by bacteriophage T1 of DNA packaging at a site between the P and Q genes of bacteriophage X, J.

Virol. PubMed Google Scholar Drexler, H., and Christensen, J. R.,Genetic crosses between restricted and unrestricted phage Ti .