期刊
NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS
卷 23, 期 6-7, 页码 891-893出版社
MARCEL DEKKER INC
DOI: 10.1081/NCN-200026037
关键词
aptamer; SELEX; pro-urokinase
Single-chain pro-urokinase is an inactive proenzyme form of human urokinase (urinary plasminogen activator) with a Mr of 50,000 which is converted to the active two-chain form by catalytic amounts of plasmin. It is used for thrombolytic therapy of acute myocardial infarction and acute ischemic stroke. We have isolated single-stranded DNA molecules with significantly increased binding affinity for human prourokinase by SELEX (systematic evolution of ligands by exponential enrichment) procedure from a pool of 1015 molecules containing 24 randomized positions which are flanked by defined regions. ssDNA from this library was hybridized with helper much less thanfixturemuch greater than, thus allowing the central random chain to fold into complex three-dimentional shapes. Sequencing data from pro-urokinase aptamers obtained after 12 selection cycles displayed a highly conserved 12-14 base region.
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