A novel means of asymmetric linker attachment to DNA molecule by RecA-mediated multistranded DNA formation.
We have developed a novel asymmetric linker attachment technology that utilizes multistranded DNA formation mediated by the RecA protein and Exonuclease I. Multistranded DNA can readily be formed at the terminus of double-stranded DNA by a complementary oligonucleotide in the presence of RecA and Exonuclease I. We have explored the possibility of applying this finding to the asymmetric attachment of linkers to specific DNA termini. We show that these unique properties of the terminal triple-stranded structure can be applied to directly clone specific DNA sequences.
Shigemori
Yasushi
Y
Kazusa DNA Research Institute, 2-6-7 Kazusa-kamatari, Kisarazu, Chiba 292-0818, Japan. syasushi@kazusa.or.jp
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry; D001412:Bacillus subtilis / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000737:chemistry*; D004269:DNA, Bacterial / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005090:Exodeoxyribonucleases / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011938:Rec A Recombinases / Q000378:metabolism*
D004269:DNA, Bacterial; D009838:Oligodeoxyribonucleotides; C051802:triplex DNA; C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D004247:DNA; D011938:Rec A Recombinases; D005090:Exodeoxyribonucleases; C028450:exodeoxyribonuclease I; C055654:endodeoxyribonuclease NruI; C047020:endodeoxyribonuclease ScaI; D015252:Deoxyribonucleases, Type II Site-Specific