In vitro selection and characterization of cellulose-binding DNA aptamers.
In vitro selection and characterization of cellulose-binding DNA aptamers.
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DOI：
10.1093/nar/gkm708
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发表时间：
2007
影响因子：
14.9
通讯作者：
Breaker, Ronald R
中科院分区：
文献类型：
--
作者：
Boese, Benjamin J;Breaker, Ronald R
许多核酸酶和适配子具有模块化的结构，使它们在与其他核苷酸序列结合时保持其功能。这种模块化功能有助于设计具有更复杂功能的RNA和DNA。我们试图创造与纤维素结合的新DNA适配子，以提供固定DNA的模块。纤维素已被用于从涂料和薄膜到药物制剂的各种应用，因此与纤维素结合的DNA适配子可能使新的应用成为可能。我们使用体外选择从随机序列DNA池中分离适体，并对两个不同的克隆进行额外的几轮突变和选择。选择一个适配子(CELAPT 14)进行序列最小化和更详细的生化分析。CELAPT 14适配子变体表现出与纤维素粉和纸张的牢固结合。此外，还设计了一种变构适配子结构，在纸层析过程中显示了ATP介导的纤维素结合。
Many nucleic acid enzymes and aptamers have modular architectures that allow them to retain their functions when combined with other nucleotide sequences. This modular function facilitates the engineering of RNAs and DNAs that have more complex functions. We sought to create new DNA aptamers that bind cellulose to provide a module for immobilizing DNAs. Cellulose has been used in a variety of applications ranging from coatings and films to pharmaceutical preparations, and therefore DNA aptamers that bind cellulose might enable new applications. We used in vitro selection to isolate aptamers from a pool of random-sequence DNAs and subjected two distinct clones to additional rounds of mutagenesis and selection. One aptamer (CELAPT 14) was chosen for sequence minimization and more detailed biochemical analysis. CELAPT 14 aptamer variants exhibit robust binding both to cellulose powder and paper. Also, an allosteric aptamer construct was engineered that exhibits ATP-mediated cellulose binding during paper chromatography.