Genetic recombination without using either restriction enzyme or PCR.
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.
Zhou
Jing-Min
JM
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Yamamoto
Yoji
Y
Uehara
Akihiko
A
Komiyama
Makoto
M
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
D004274:DNA, Recombinant; D020135:Peptide Nucleic Acids; D049452:Green Fluorescent Proteins; D004262:DNA Restriction Enzymes; D015719:Single-Strand Specific DNA and RNA Endonucleases