期刊
CHEMISTRY-A EUROPEAN JOURNAL
卷 20, 期 9, 页码 2420-2424出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/chem.201304292
关键词
aptamers; biosensors; molecular competition; protein detection; rolling circle amplification
资金
- Natural Sciences and Engineering Research Council of Canada (NSERC)
- SENTINEL Bioactive Paper Network
- National Basic Research Program of China [2011CB935704]
- National Natural Science Foundation of China [21235004, 21327806]
- European Union Seventh Framework Programme [260600]
- China Scholarship Council
- NSERC
Certain DNA polymerases, such as phi 29 DNA polymerase, can isothermally copy the sequence of a circular template round by round in a process known as rolling circle amplification (RCA), which results in super-long single-stranded (ss) DNA molecules made of tandem repeats. The power of RCA reflects the high processivity and the strand-displacement ability of these polymerases. In this work, the ability of phi 29DNAP to carry out RCA over circular templates containing a protein-binding DNA aptamer sequence was investigated. It was found that protein-aptamer interactions can prevent this DNA polymerase from reading through the aptameric domain. This finding indicates that protein-binding DNA aptamers can form highly stable complexes with their targets in solution. This novel observation was exploited by translating RCA arrest into a simple and convenient colorimetric assay for the detection of specific protein targets, which continues to showcase the versatility of aptamers as molecular recognition elements for biosensing applications.
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