Improvement of the dideoxy chain termination method of DNA sequencing by use of deoxy-7-deazaguanosine triphosphate in place of dGTP

Mizusawa, Saeko; Nishimura, Susumu; Seela, Frank

doi:10.1093/nar/14.3.1319

Cited by 698 publications

(307 citation statements)

References 6 publications

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“…The DNA sequence was determined by the dideoxynucleotide method of Sanger et al (1977). using (alphaS 35 )dATP (Amersham) and 7-deaza-dGTP (Boehringer) instead of dGTP, since this results in better resolution of GC-rich regions in polyacrylamide gels (Mizusawa et al , 1986). Sequence analysis was performed using the facilities of the Unite d'lnformatique Scientifique of the Pasteur Institute.…”

Section: Methodsmentioning

confidence: 99%

The calmodulin‐sensitive adenylate cyclase of Bordetella pertussis: cloning and expression in Escherichia col

Glaser

Ladant

Sezer

et al. 1988

Molecular Microbiology

306

172

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SummaryThe adenylate cyclase toxin of the prokaryote Bordetella pertussis is stimulated by the eukaryotic regulatory protein, calmodulin. A general strategy, using the adenylate-cyclase-calmodulin interaction as a tool, has permitted cloning and expression of the toxin in Escherichia coli in the absence of any B. pertussis trans-activating factor. We show that the protein is synthesized in a large precursor form composed of 1706 amino acids. The calmodulin-stimulated catalytic activity resides in the amino-terminal 450 amino acids of the adenylate cyclase. The enzyme expressed in E. coli is recognized in Western blots by antibodies directed against purified B. pertussis adenylate cyclase, and its activity is inhibited by these antibodies.

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Section: Methodsmentioning

confidence: 99%

The calmodulin‐sensitive adenylate cyclase of Bordetella pertussis: cloning and expression in Escherichia col

Glaser

Ladant

Sezer

et al. 1988

Molecular Microbiology

306

172

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“…Nucleotide sequences of overlapping clones of this insert were determined by the method of Chen and Seeburg (1985). Nucleotide sequences in both directions of overlapping deletion clones were determined by the procedure of Sanger et al (1977), using 7-deaza dGTP to clarify regions previously obscured by GC compression (Mizusawa et al, 1986). Sequence data were compiled and comparative analyses were made using the GCG program (University of Wisconsin, Madison), the BLASTN, BLASTX, and BESTFIT programs developed by the National Centre for Biotechnology Information (Altschul et al, 1990;Devereux et al, 1984;Gish and States, 1993), and the GENECOMPAR program (Applied Maths Department, Kortrijk, Belgium).…”

Section: Dna Preparationmentioning

confidence: 99%

**Characterization of flagella genes of Agrobacterium tumefaciens, and the effect of a bald strain on virulence**

Chesnokova

Coutinho

Khan

et al. 1997

Molecular Microbiology

104

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Because the 54 promoter is associated with genes regulated by physiological changes in various bacteria, the flaC gene might be similarly regulated in response to A. tumefaciens responding to host plant stimuli. Virulence studies showed that the bald strain was consistently reduced in virulence below that of the parental wild-type strain by at least 38%. The difference is statistically significant and suggests that the flagella may play a role in facilitating virulence.

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“…Both ~X -[~~P ]~A T P (Amersham) and ~X -[~~S ]~A T P (New England Nuclear) were used as radioactive tracer in different experiments. Aberrations in the banding pattern were relieved by using 'sequenase' (United States Biochemicals) reactions with either dITP (Mills & Kramer, 1979) or 7-deaza-dGTP (Mizusawa et al, 1986) in place of dGTP.…”

Section: Methodsmentioning

confidence: 99%

Discovery of an insertion sequence, IS116, from Streptomyces clavuligerus and its relatedness to other transposable elements from actinomycetes

Leskiw

Mevarech

Barritt

et al. 1990

Journal of General Microbiology

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We have identified an insertion sequence, IS116, present in Streptomyces clavuligerus at one copy per genome. The element was discovered as a 1.4 kb insertion into the multicopy plasmid pIJ702 after propagation in S. clavuligerus. The nucleotide sequence of IS116 and the flanking sequences from pIJ702 have been determined. The junctions with pIJ702 show no target site duplication and there are no inverted repeats at the ends of the element. One putative coding open reading frame of 1197 bp was identified which would code for a protein product of 399 amino acids. This protein resembles deduced integrase/transposase proteins specified by three other transposable elements of actinomycetes: IS110 and the mini-circle from Stveptomyces coelicolor A3(2), and -most particularly -IS900 of Mycobacterium paratuberculosis. Two regions that are relatively conserved among these gene products show features found in similar positions in many reverse transcriptases. IS116 and IS900 are also closely similar in their general organization and (apparently) in their insertion site specificity, whereas IS110 and the mini-circle are quite different in these features.

show abstract

Improvement of the dideoxy chain termination method of DNA sequencing by use of deoxy-7-deazaguanosine triphosphate in place of dGTP

Cited by 698 publications

References 6 publications

The calmodulin‐sensitive adenylate cyclase of Bordetella pertussis: cloning and expression in Escherichia col

The calmodulin‐sensitive adenylate cyclase of Bordetella pertussis: cloning and expression in Escherichia col

**Characterization of flagella genes of Agrobacterium tumefaciens, and the effect of a bald strain on virulence**

Discovery of an insertion sequence, IS116, from Streptomyces clavuligerus and its relatedness to other transposable elements from actinomycetes

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