Rapid and easy sequencing of large linear double-stranded DNA and supercoiled plasmid DNA

Zagursky, Robert J.; Baumeister, Kirk; Lomax, N B; Berman, Michael L.

doi:10.1016/0735-0651(85)90011-1

Cited by 441 publications

(174 citation statements)

References 12 publications

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“…The latter were then subjected to DNA sequence analysis. The tRNA region of the pZ189 vector was sequenced using a modification of the Sanger dideoxyribonucleotide procedure which allows sequence determination directly from the double-stranded plasmid DNA (25). The possibility that mutations arise in the bacterial rather than the mammalian host cell as a result of a low level of persistent lesions formed in pZ189 DNA during H 2 O 2 treatment of the transfected cells is difficult to exclude experimentally.…”

Section: Mutant Selection and Characterizationmentioning

confidence: 99%

Mutagenesis by hydrogen peroxide treatment of mammalian cells: a molecular analysis

Moraes¹,

Keyse²,

Tyrrell³

1990

Carcinogenesis

113

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Hydrogen peroxide is an oxidizing agent which can be generated intracellularly either during normal metabolism or by treatment with external agents including solar UV radiation. Simian cells (CV-1) transfected with the SV40-based shuttle vector plasmid pZ189 have been treated with H 2 O 2 and then incubated to allow repair and replication of the plasmid. The frequency of mutations at the supF locus of the recovered plasmid increases by a factor of up to four over the spontaneous value. The nucleotide changes associated with 100 spontaneous and 100 H 2 O 2 -induced mutants have been determined directly by sequencing a 150 bp fragment that includes the entire supF tRNA coding region. Deletions were observed in -45% of both the spontaneous and induced mutants, whereas single or multiple base changes arose in 68 and 57% of the induced and spontaneous mutants respectively. The spectrum of induced mutations is characterized by (i) the occurrence of deletions associated with base changes (16% of all mutants analysed) and (ii) small deletions of 3 bp and less (51% of all deletion mutants sequenced). Sixty-five per cent (15 out of 23) of all small deletions (spontaneous and induced) are associated with runs of between two and five identical bases and eight of them arise at a mutational 'hotspot' region of five cytosines between bp 172 and 176. The majority (19 out of 30) of completely sequenced deletions observed in the spontaneous spectrum contain either (i) small (2-10 bp) direct repeat sequences that lie immediately outside one deletion terminus and immediately inside the second deletion terminus or (ii) small (2-3 bp) inverted repeat sequences lying immediately inside the two deletion termini. Most deletions that we have observed are therefore likely to arise as a consequence of specific aspects of DNA structure. IntroductionOxygen radicals appear to be involved in both the ageing process and in the initiation and progression of many human diseases, including cancer. A knowledge of the nature of mutations arising in DNA as a result of oxygen radical attack should provide important clues to understanding the molecular events that underlie these pathological changes. The characterization of the genotoxic action of the oxidizing agent, hydrogen peroxide, is particularly relevant in this respect for several reasons. In addition

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Section: Mutant Selection and Characterizationmentioning

confidence: 99%

Mutagenesis by hydrogen peroxide treatment of mammalian cells: a molecular analysis

Moraes¹,

Keyse²,

Tyrrell³

1990

Carcinogenesis

113

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“…]) was done as in [12,13]. LW-AN32 was derived from aa 13 to 22 of the published sequence [6], LW-21 and LW-77 were derived from our nt sequences and LW-31 was derived from the sequence of exon 14 [9].…”

Section: Introduction 2 Materials and Methodsmentioning

confidence: 99%

cDNA clones completing the nucleotide and derived amino acid sequence of the alpha 1 chain of basement membrane (type IV) collagen from mouse

Wood

Theriault²,

Vögeli

1988

FEBS Letters

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Six cDNA clones add 3549 nucleotides to the DNA sequence of the alpha 1 chain of basement membrane (type IV) collagen. Thus the complete nucleotide and derived amino acid sequence of the alpha 1 type IV collagen with 5007 nucleotides coding for 1669 amino acids with a calculated M r of 160 827 is known. The six cDNA clones cover the putative N‐terminal signal peptide, the 7 S region and two thirds of the helical region extending into the previously published murine nucleotide sequence [(1986) Gene 43, 301]. The protein sequence for 289 amino acids of the helical region adjacent to the 7 S region has not previously been published for any species.

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“…Sequencing of the plasmid DNA was carried out with (5'-P)-labeled oligonucleotides, T3 and T7, according to Zagursky et al [20].…”

Section: Dna Sequencingmentioning

confidence: 99%

Metribuzin resistance in photoautotrophic Chenopodium rubrum cell cultures

et al. 1993

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Rapid and easy sequencing of large linear double-stranded DNA and supercoiled plasmid DNA

Cited by 441 publications

References 12 publications

Mutagenesis by hydrogen peroxide treatment of mammalian cells: a molecular analysis

Mutagenesis by hydrogen peroxide treatment of mammalian cells: a molecular analysis

cDNA clones completing the nucleotide and derived amino acid sequence of the alpha 1 chain of basement membrane (type IV) collagen from mouse

Metribuzin resistance in photoautotrophic Chenopodium rubrum cell cultures

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