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PubMed · 17150732

Genetic recombination without using either restriction enzyme or PCR.

Jing-Min Zhou·Yoji Yamamoto·Akihiko Uehara·Makoto Komiyama
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Abstract

We recently prepared artificial restriction DNA cutters (ARCUT) for site-selective scission of double-stranded DNA by combining Ce(IV)/EDTA complex with a pair of pseudo-complementary peptide nucleic acids (pcPNAs). Here we report an improved method for genetic recombination using ARCUT. The key point is to treat the scission fragments with nuclease S1 (a single-stranded DNA specific enzyme) and form blunt ends. By this procedure, these scission fragments are efficiently ligated with foreign DNA fragments having blunt ends, providing desired recombinant DNA in high yields. Neither restriction enzyme nor PCR amplification is required.

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DNA Restriction EnzymesDNA, RecombinantGreen Fluorescent ProteinsPeptide Nucleic AcidsPolymerase Chain ReactionProtein EngineeringRecombination, GeneticSingle-Strand Specific DNA and RNA EndonucleasesJournal ArticleResearch Support, Non-U.S. Gov't

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Jing-Min Zhou, Yoji Yamamoto, Akihiko Uehara, Makoto Komiyama. 2005. Genetic recombination without using either restriction enzyme or PCR.. https://doi.org/10.1093/nass%2F49.1.259

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