2020
DOI: 10.1002/cpnc.121
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Mutation Analysis of L‐Thymidine‐Induced Replication Products Using a Restriction Enzyme–Mediated Assay

Abstract: This article describes experimental and analytical procedures for evaluating the efficiency and fidelity of DNA replication containing mirror‐image thymidine (L‐T) in E. coli. The procedure involves construction of DNA recombinants containing a restriction enzyme (PstI) recognition site in which the L‐T lesion is site‐specifically located within the PstI recognition sequence (CTGCAG). The recombinants are transfected into DH5α cells. DNA is extracted, amplified, and cleaved into relatively short fragments usin… Show more

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Cited by 2 publications
(10 citation statements)
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“…l-dG→T, dG, dC and dA. Thus, by monitoring the products and evaluating bypass e ciency and mutation frequency from the replication of l-nucleosides-containing strand, we were able to study the effect of lnucleosides on DNA replication in vitro and in cells 20,21 .…”
Section: Resultsmentioning
confidence: 99%
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“…l-dG→T, dG, dC and dA. Thus, by monitoring the products and evaluating bypass e ciency and mutation frequency from the replication of l-nucleosides-containing strand, we were able to study the effect of lnucleosides on DNA replication in vitro and in cells 20,21 .…”
Section: Resultsmentioning
confidence: 99%
“…We examined previously the cytotoxic and mutagenic properties of l-T in E. coli as well as several DNA polymerases 20,21 , which revealed distinctly different bypass e ciencies and mutagenic properties in different replication systems. This study was systematic interrogation about the impact of the lnucleosides on the bypass e ciency and delity of DNA replication.…”
Section: Discussionmentioning
confidence: 99%
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