Aptamer beacons for the direct detection of proteins

被引:482
作者
Hamaguchi, N
Ellington, A
Stanton, M
机构
[1] Brandeis Univ, Rosenstiel Basic Med Sci Res Ctr, Waltham, MA 02454 USA
[2] Univ Texas, MBB 1 206, ICMB A4800, Austin, TX 78712 USA
关键词
proteins; analysis; fluorescent dyes; chemistry; chemical synthesis; DNA; single stranded; nucleic acid conformation; ligands;
D O I
10.1006/abio.2001.5169
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We have designed a new class of molecules, which we term aptamer beacons, for detecting a wide range of ligands. Similar to molecular beacons, aptamer beacons can adopt two or more conformations, one of which allows ligand binding. A fluorescence-quenching pair is used to report changes in conformation induced by ligand binding. An anti-thrombin aptamer was engineered into an aptamer beacon by adding nucleotides to the 5 ' -end which are complementary to nucleotides at the 3 ' -end of the aptamer. In the absence of thrombin, the added nucleotides will form a duplex with the 3 ' -end, forcing the aptamer beacon into a stem-loop structure. In the presence of thrombin, the aptamer beacon forms the ligand-binding structure. This conformational change causesa change in the distance between a fluorophore attached to the 5 ' -end and a quencher attached to the 3 ' -end. Aptamer beacon can be a sensitive tool for detecting proteins and other chemical compounds. (C) 2001 Academic Press.
引用
收藏
页码:126 / 131
页数:6
相关论文
共 31 条
  • [1] Nucleic acid selection as a tool for drug discovery
    Bacher, JM
    Ellington, AD
    [J]. DRUG DISCOVERY TODAY, 1998, 3 (06) : 265 - 273
  • [2] SELECTION OF SINGLE-STRANDED-DNA MOLECULES THAT BIND AND INHIBIT HUMAN THROMBIN
    BOCK, LC
    GRIFFIN, LC
    LATHAM, JA
    VERMAAS, EH
    TOOLE, JJ
    [J]. NATURE, 1992, 355 (6360) : 564 - 566
  • [3] Automated RNA selection
    Cox, JC
    Rudolph, P
    Ellington, AD
    [J]. BIOTECHNOLOGY PROGRESS, 1998, 14 (06) : 845 - 850
  • [4] Crameri R, 1999, COMB CHEM HIGH T SCR, V2, P63
  • [5] Famulok M, 1999, CURR TOP MICROBIOL, V243, P123
  • [6] Using molecular beacons to probe molecular interactions between lactate dehydrogenase and single-stranded DNA
    Fang, XH
    Li, JJ
    Tan, WH
    [J]. ANALYTICAL CHEMISTRY, 2000, 72 (14) : 3280 - 3285
  • [7] GOLD L, 1995, NUCL ACIDS S SER, V33, P20
  • [8] The use of recombinant antibodies in proteomics
    Holt, LJ
    Enever, C
    de Wildt, RMT
    Tomlinson, IM
    [J]. CURRENT OPINION IN BIOTECHNOLOGY, 2000, 11 (05) : 445 - 449
  • [9] Jayasena SD, 1999, CLIN CHEM, V45, P1628
  • [10] In vitro selection of signaling aptamers
    Jhaveri, S
    Rajendran, M
    Ellington, AD
    [J]. NATURE BIOTECHNOLOGY, 2000, 18 (12) : 1293 - 1297