title: Genetic recombination without using either restriction enzyme or PCR.
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.
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
authors.forename: Jing-Min
authors.initials: JM
authors.lastname: Zhou
authors.forename: Yoji
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Akihiko
authors.initials: A
authors.lastname: Uehara
authors.forename: Makoto
authors.initials: M
authors.lastname: Komiyama
mesh_terms: D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000235:genetics; D020135:Peptide Nucleic Acids / Q000737:chemistry; D016133:Polymerase Chain Reaction; D015202:Protein Engineering / Q000379:methods*; D011995:Recombination, Genetic; D015719:Single-Strand Specific DNA and RNA Endonucleases
chemical_list: D004274:DNA, Recombinant; D020135:Peptide Nucleic Acids; D049452:Green Fluorescent Proteins; D004262:DNA Restriction Enzymes; D015719:Single-Strand Specific DNA and RNA Endonucleases