A Fok I/DNA machine that duplicates its analyte gene sequence.
A method for the amplified analysis of a gene sequence by a Fok I/DNA machine is described. A tailored nucleic acid that includes a recognition sequence for the analyzed gene and a hybridized sequence of the gene itself acting as the "fuel" substrate of the "machine". Upon recognition of the gene by the "fuel" substrate, the autonomous Fok I-induced cleavage of the resulting duplex proceeds, resulting in the duplication of the gene sequence and the horseradish peroxidase mimicking hemin/G-quadruplex DNAzyme as a chemiluminescence reporter. The autonomous duplication of the gene sequence and the catalytic activity of the DNAzyme provide a double amplification path that enables the analysis of target DNA with a detection limit of 1 x 10(-14) M.
Weizmann
Yossi
Y
Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Cheglakov
Zoya
Z
Willner
Itamar
I
D002384:Catalysis; D004247:DNA / Q000737:chemistry*; D021881:DNA, Catalytic / Q000737:chemistry; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry*; D054856:G-Quadruplexes; D017930:Genes, Reporter; D006427:Hemin / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000737:chemistry; D008165:Luminol / Q000737:chemistry; D008956:Models, Chemical; D008957:Models, Genetic; D009690:Nucleic Acid Conformation; D010100:Oxygen / Q000737:chemistry; D017422:Sequence Analysis, DNA; D013997:Time Factors
D021881:DNA, Catalytic; D008165:Luminol; D006427:Hemin; D004247:DNA; D006861:Hydrogen Peroxide; C035042:endodeoxyribonuclease FokI; D015252:Deoxyribonucleases, Type II Site-Specific; D010100:Oxygen