Physical mapping of bacteriophage T4

Niggemann, Elisabeth; Green, Ian; Meyer, Hans‐Peter; Rüger, Wolfgang

doi:10.1007/bf00272920

Cited by 17 publications

(5 citation statements)

References 38 publications

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“…Identification of genes 55, o!gt, 47 and 46 On the basis of mapping data (Wood and Revel, 1976;Niggemann et al, 1981;Spicer and Konigsberg, 1983) genes aogt, 47, 46, and possibly gene 55, were expected to be found within the DNA region studied here. To identify these genes, selected subclones used during the sequencing procedure were subjected to marker rescue tests.…”

Section: Resultsmentioning

confidence: 80%

“…Determination of the DNA sequence The starting material for this investigation was cytosinecontaining T4 DNA isolated from purified phage particles of strain T4 JSW800. Since the physical map of T4 is now well established and aligned with the genetic map (Niggemann et al, 1981;Kutter and Ruger, 1983), the restriction fragments spanning the DNA region which we intended to study, could be isolated from the mass of total T4 DNA. With the exception of a 2.3-kb XhoI-Sall fragment (map position 36.1 -33.8 kb) which we did not succeed in cloning in its entirety, the restriction fragments were first amplified in vector DNA: a BgllI-XhoI fragment (map position 39.4-37.0 kb) and a XbaI-ClaI fragment (map position 34.3 -32.75 kb) in M13mp8, and two HindIIl fragments (map positions 38.55-35.85 kb and 33.7-32.3 kb) in pBR322.…”

Section: Resultsmentioning

confidence: 99%

“…Enzymes T4 ligase was isolated by a combination of methods describd in the literature from a ligase-overproducing E. coli strain (Wilson and Murray, 1979;Murray et al, 1979;Knopf, 1977;Tait et al, 1980). Restriction endonucleases were either isolated as described (Niggemann et al, 1981) or they were purchased from Boehringer or Bethesda Research Laboratories. E. coli DNA polymerase I (Klenow fragment), nuclease S1 and alkaline phosphatase were obtained from Boehringer, Mannheim.…”

Section: Methodsmentioning

confidence: 99%

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Genes 55, alpha gt, 47 and 46 of bacteriophage T4: the genomic organization as deduced by sequence analysis.

Gram¹,

Rüger²

1985

The EMBO Journal

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The nucleotide sequence of T4 genes 55, alpha gt, 47 and 46 was determined by a combination of ‘classical’ procedures and a shotgun approach. Small DNA fragments generated by frequent cleavage with restriction enzymes or by sonication of restriction fragments were cloned in phage M13 vectors and sequenced by the dideoxy method. The positions of the genes were determined by marker rescue between the corresponding T4 amber mutants and the cloned T4 DNA fragments used in the sequencing experiments. The sequence gives an insight into the organization of this 7.1‐kb early region of the T4 genome and shows that genetically ‘silent’ portions within this region are not void of genetic information.

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Section: Resultsmentioning

confidence: 80%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Genes 55, alpha gt, 47 and 46 of bacteriophage T4: the genomic organization as deduced by sequence analysis.

Gram¹,

Rüger²

1985

The EMBO Journal

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“…A 15 kb SnaI fragment was isolated from digested JW800 DNA by phenol extraction (Niggemann et al, 1981) of DNA from pieces cut out of 0.5% agarose gels. The SmiaI fragment was further digested by SphI and ClaI endonucleases and fragments were cloned into Ml3mpl9.…”

Section: Sequencing Strategymentioning

confidence: 99%

The bacteriophage T4 gene for the small subunit of ribonucleotide reductase contains an intron.

Sjoberg¹,

Hahne²,

Mathews³

et al. 1986

The EMBO Journal

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The bacteriophage T4 gene nrdB codes for the small subunit of the enzyme ribonucleotide reductase. The T4 nrdB gene was localized between 136.1 kb and 137.8 kb in the T4 genetic map according to the deduced structural homology of the protein to the amino acid sequence of its bacterial counterpart, the B2 subunit of Escherichia coli. This positions the C‐terminal end of the T4 nrdB gene approximately 2 kb closer to the T4 gene 63 than earlier anticipated from genetic recombinational analyses. The most surprising feature of the T4 nrdB gene is the presence of an approximately 625 bp intron which divides the structural gene into two parts. This is the second example of a prokaryotic structural gene with an intron. The first prokaryotic intron was reported in the nearby td gene, coding for the bacteriophage T4‐specific thymidylate synthase enzyme. The nucleotide sequence at the exon‐intron junctions of the T4 nrdB gene is similar to that of the junctions of the T4 td gene: the anticipated exon‐intron boundary at the donor site ends with a TAA stop codon and there is an ATG start codon at the putative downstream intron‐exon boundary of the acceptor site. In the course of this work the denA gene of T4 (endonuclease II) was also located.

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“…In E. coli, any defined sequence of given length would be equally probable, on the average. In T4 DNA, a sequence containing predominantly G * C pairs would be, on the average, less frequent than a sequence of the same length containing predominantly A * T pairs, an expectation borne out by several studies of the occurrence of various restriction sites in T4 DNA (9,10,33,39,40), and also less frequent, per unit length, in T4 DNA than in E. coli DNA. It is perhaps not surprising that the host DNA from cells infected with T4 42-46-56-phage yielded only a broad smear with no evidence of discretely sized fragments, since the E. coli chromosome is about 20 times more complex than that of T4.…”

Section: Discussionmentioning

confidence: 98%

In vivo cleavage of cytosine-containing bacteriophage T4 DNA to genetically distinct, discretely sized fragments

Carlson¹,

Wiberg²

1983

J Virol

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Source" and reference 42 amN55 x 5 dCMP hydroxymethylase-This laboratory 42 amN122 x 5 dCMP hydroxymethylase-This laboratory 46 amB14 x 5 46-47 nuclease-This laboratory 46 amN126 46-47 nuclease-This laboratory 56 amE51 x 5 dCTPase-; Cyt This laboratory 56 amE114 x 5 dCTPase-; Cyt This laboratory denA S112 x 5 endonuclease 11-This laboratory (19) denA SP4 x 5 endonuclease lI-This laboratory (19) denB D2a2 x 5 endonuclease IV-Vetter and Sadowski (51) denB D2a23 x 5 endonuclease IV-Vetter and Sadowski (51) denB saA&9 endonuclease IV-(12)

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Physical mapping of bacteriophage T4

Cited by 17 publications

References 38 publications

Genes 55, alpha gt, 47 and 46 of bacteriophage T4: the genomic organization as deduced by sequence analysis.

Genes 55, alpha gt, 47 and 46 of bacteriophage T4: the genomic organization as deduced by sequence analysis.

The bacteriophage T4 gene for the small subunit of ribonucleotide reductase contains an intron.

In vivo cleavage of cytosine-containing bacteriophage T4 DNA to genetically distinct, discretely sized fragments

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