Mutation spectra of M13 vectors containing site-specific cis-syn, trans-syn-I, (6-4), and dewar pyrimidone photoproducts of thymidylyl-(3′→5′)-thymidine in Escherichia coli under SOS conditions

Colin A. Smith, Mu Wang, Nan Jiang, Linda Che, Xiaodong Zhao, John Stephen Taylor

Research output: Contribution to journalArticle

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Abstract

The mutation spectra of cis-syn, trans-syn-I, (6-4), and Dewar pyrimidone photoproducts of the TT site of AATTAA and TATTAT in the (-) strand of a heteroduplex M13 vector were obtained in an excision and photoreversal repair deficient Escherichia coli host under SOS conditions. Oligonucleotides containing site-specific photoproducts were annealed to a complementary uracil-containing (+) strand that contained one or more unique pairs of nucleotide mismatches and used to prime (-) strand synthesis with a DNA polymerase and dNTPs. Following DNA synthesis, the reaction mixtures were incubated with T4 DNA ligase and ATP and then used to transfect SOS-induced competent CSRO6F′ cells (uvrA6 and phr-1). The transfectants were plated, gridded, and probed by oligonucleotides specific for progeny of the (-) and (+) strands. Individual progeny of the photoproduct-containing (-) strands were plaque purified and sequenced by the dideoxy method. The cis-syn and trans-syn-I dimers were found not to be very mutagenic (

Original languageEnglish (US)
Pages (from-to)4146-4154
Number of pages9
JournalBiochemistry
Volume35
Issue number13
StatePublished - Apr 2 1996
Externally publishedYes

Fingerprint

Pyrimidinones
Dewars
Oligonucleotides
Thymidine
Escherichia coli
DNA Ligases
Mutation
Uracil
DNA-Directed DNA Polymerase
DNA Repair
Dimers
Repair
Nucleotides
Adenosine Triphosphate
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mutation spectra of M13 vectors containing site-specific cis-syn, trans-syn-I, (6-4), and dewar pyrimidone photoproducts of thymidylyl-(3′→5′)-thymidine in Escherichia coli under SOS conditions. / Smith, Colin A.; Wang, Mu; Jiang, Nan; Che, Linda; Zhao, Xiaodong; Taylor, John Stephen.

In: Biochemistry, Vol. 35, No. 13, 02.04.1996, p. 4146-4154.

Research output: Contribution to journalArticle

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abstract = "The mutation spectra of cis-syn, trans-syn-I, (6-4), and Dewar pyrimidone photoproducts of the TT site of AATTAA and TATTAT in the (-) strand of a heteroduplex M13 vector were obtained in an excision and photoreversal repair deficient Escherichia coli host under SOS conditions. Oligonucleotides containing site-specific photoproducts were annealed to a complementary uracil-containing (+) strand that contained one or more unique pairs of nucleotide mismatches and used to prime (-) strand synthesis with a DNA polymerase and dNTPs. Following DNA synthesis, the reaction mixtures were incubated with T4 DNA ligase and ATP and then used to transfect SOS-induced competent CSRO6F′ cells (uvrA6 and phr-1). The transfectants were plated, gridded, and probed by oligonucleotides specific for progeny of the (-) and (+) strands. Individual progeny of the photoproduct-containing (-) strands were plaque purified and sequenced by the dideoxy method. The cis-syn and trans-syn-I dimers were found not to be very mutagenic (",
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AU - Che, Linda

AU - Zhao, Xiaodong

AU - Taylor, John Stephen

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