Using a deoxyribozyme ligase and rolling circle amplification to detect a non-nucleic acid analyte, ATP.
An allosteric ribozyme (aptazyme) has been used to transduce the binding of a small organic analyte (ATP) into the ligation of a circular template for rolling circle amplification (RCA). An ATP-activated deoxyribozyme ligase was immobilized on a glass slide and, upon addition of ATP, catalyzed the ligation of a circular padlock probe. The ligated products could be directly amplified and visualized via RCA. The coupled reaction exhibited could detect as little as 1 muM of ATP and could discriminate against structurally similar nucleotides such as GTP, CTP, and UTP. Cooperative ATP activation of the deoxyribozyme was faithfully mimicked by RCA, yielding an amplified "switch" that was responsive to ATP concentration.
Cho
Eun Jeong
EJ
Department of Chemistry and Biochemistry, Institute for Cell and Molecular Biology, University of Texas at Austin, Austin, Texas 78712, USA.
Yang
Litao
L
Levy
Matthew
M
Ellington
Andrew D
AD
D000255:Adenosine Triphosphate / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D011088:DNA Ligases / Q000737:chemistry* / Q000378:metabolism; D021881:DNA, Catalytic / Q000737:chemistry* / Q000378:metabolism; D004270:DNA, Circular / Q000737:chemistry / Q000378:metabolism; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009690:Nucleic Acid Conformation
D021881:DNA, Catalytic; D004270:DNA, Circular; D000255:Adenosine Triphosphate; D004247:DNA; D011088:DNA Ligases