Identifying the Location of a Single Protein along the DNA Strand Using Solid-State Nanopores
| Autor: | Duyen Thi Ngoc Huynh, Jae-Seok Yu, Min-Cheol Lim, Ki-Bum Kim, Young-Rok Kim, Hyun-Mi Kim, Hyung-Jun Kim |
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| Rok vydání: | 2015 |
| Předmět: |
Models
Molecular Zinc finger Base Sequence HMG-box General Engineering General Physics and Astronomy RNA Zinc Fingers DNA Plasma protein binding Biology DNA-binding protein Molecular biology DNA-Binding Proteins Nanopores chemistry.chemical_compound Nanopore Electricity chemistry Biophysics Nucleic Acid Conformation General Materials Science Binding site |
| Zdroj: | ACS Nano. 9:5289-5298 |
| ISSN: | 1936-086X 1936-0851 |
| DOI: | 10.1021/acsnano.5b00784 |
| Popis: | Solid-state nanopore has been widely studied as an effective tool to detect and analyze small biomolecules, such as DNA, RNA, and proteins, at a single molecule level. In this study, we demonstrate a rapid identification of the location of zinc finger protein (ZFP), which is bound to a specific locus along the length of a double-stranded DNA (dsDNA) to a single protein resolution using a low noise solid-state nanopore. When ZFP labeled DNAs were driven through a nanopore by an externally applied electric field, characteristic ionic current signals arising from the passage of the DNA/ZFP complex and bare DNA were detected, which enabled us to identify the locations of ZFP binding site. We examined two DNAs with ZFP binding sites at different positions and found that the location of the additional current drop derived from the DNA/ZFP complex is well-matched with a theoretical one along the length of the DNA molecule. These results suggest that the protein binding site on DNA can be mapped or that genetic information can be read at a single molecule level using solid-state nanopores. |
| Databáze: | OpenAIRE |
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