SELEX for tubulin affords specific T-rich DNA aptamers. Systematic evolution of ligands by exponeential enrichment.
We succeeded in acquiring two DNA aptamers that selectively recognize tubulin by the SELEX method. A pool of single-stranded oligo-DNAs including a random region of 59 nucleotides was screened by SELEX for tubulin purified from calf-brain as a target. After 20 repetitions of selection round, the library converged on specific T-rich sequences. The binding activity of T-rich clones was analyzed by the SPR sensor to determine their dissociation constants to be in the order of 10 microM.
Fukusaki
E
E
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita 565-0871, Japan. fukasaki@bio.eng.osaka-u.ac.jp
Hasunuma
T
T
Kajiyama
S
S
Okazawa
A
A
Itoh
T J
TJ
Kobayashi
A
A
D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D008024:Ligands; D008870:Microtubules / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D017422:Sequence Analysis, DNA / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*
D004277:DNA, Single-Stranded; D008024:Ligands; D014404:Tubulin