HELIPORE
DNA sequencing using helicase-modified alpha-hemolysin nanopores
| Coordinatore | THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
Organization address
address: University Offices, Wellington Square contact info |
| Nazionalità Coordinatore | United Kingdom [UK] |
| Totale costo | 209˙033 € |
| EC contributo | 209˙033 € |
| Programma | FP7-PEOPLE
Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) |
| Code Call | FP7-PEOPLE-2011-IEF |
| Funding Scheme | MC-IEF |
| Anno di inizio | 2012 |
| Periodo (anno-mese-giorno) | 2012-04-16 - 2014-04-15 |
Partecipanti
| # | ||||
|---|---|---|---|---|
| 1 |
THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
Organization address
address: University Offices, Wellington Square contact info |
UK (OXFORD) | coordinator | 209˙033.40 |
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Obiettivo del progetto (Objective)
'Most of the current, second generation, methods for DNA sequencing are based on alternating cycles of enzyme manipulation and imaging the incorporation of fluorescently labeled nucleotides. These methods, however, hold several disadvantages, mainly the long duration of each sequencing experiment and the requirement for fluorescently labeled nucleotides and primer sequences. Tremendous efforts have been taken in order to develop cheap and ultra-fast sequencing systems. Specifically, the sequencing of single DNA molecules is of great importance, as no pre-amplification step and no labeled nucleotides are required. I intend to use biological nanometer-scale pores (alpha-hemolysin) modified with a DNA manipulating enzyme, a helicase, embedded in a membrane that separates two compartments, each contains an Ag/AgCl electrode, for the sequencing of single DNA molecules. Based on previous results, I expect that the ionic current, associated with DNA translocation through the nanopore, will exhibit a specific pattern for each nucleobase, allowing sequence determination. I intend to construct two different systems, in the first system the helicase will be soluble in the cis compartment, whereas in the second configuration the helicase will be attached to the nanopore via double stranded DNA linkers. The helicase enzyme will unwind double stranded DNA, form a ratchet-like movement of the DNA strand through the pore and control the pace of DNA translocation, leading to efficient sequencing.'