Peptide nucleic acid (PNA) facilitates multistranded hybrid formation between linear double-stranded DNA targets and RecA protein-coated complementary single-stranded DNA probes.
RecA protein-coated single-stranded DNA probes, known as RecA nucleoprotein filaments, bind specifically to homologous DNA sequences within double-stranded DNA targets, forming multistranded probe-target DNA hybrids. This DNA hybridization reaction can be used for sequence-specific gene capture, gene modification, and gene regulation. Thus, factors that enhance the efficiency of the hybridization reaction are of significant practical importance. We show here that the hybridization of a peptide nucleic acid (PNA) within or adjacent to the probe-target homology region significantly enhances the yield of hybrid DNA formed in the reaction between linear double-stranded DNA targets and RecA protein-coated complementary single-stranded (css)DNA probes. The possible mechanisms and the advantages of using RecA nucleoprotein filaments in combination with PNA for genomic DNA cloning and mutagenesis are presented.
Belotserkovskii
Boris P
BP
Pangene Corporation, 5500 Stewart Avenue, Fremont, California 94538, USA.
Zarling
David A
DA
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes; D004277:DNA, Single-Stranded / Q000737:chemistry*; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008274:Magnesium / Q000737:chemistry; D008969:Molecular Sequence Data; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D011938:Rec A Recombinases / Q000737:chemistry*
D015342:DNA Probes; D004277:DNA, Single-Stranded; D020135:Peptide Nucleic Acids; D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase; D011938:Rec A Recombinases; D008274:Magnesium