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Translocation of double stranded DNA through membrane adapted phi29 motor protein nanopore

Biological pores have been used to study the transport of DNA and other molecules but most pores have channels that allow only the movement of small molecules and single-stranded DNA and RNA. The bacteriophage phi29 DNA-packaging motor, which allows double-stranded DNA to enter and exit during a vir...

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Autores principales: Wendell, David, Jing, Peng, Geng, Jia, Subramaniam, Varuni, Lee, Tae Jin, Montemagno, Carlo, Guo, Peixuan
Formato: Texto
Lenguaje:English
Publicado: 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2777743/
https://www.ncbi.nlm.nih.gov/pubmed/19893523
http://dx.doi.org/10.1038/nnano.2009.259
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author Wendell, David
Jing, Peng
Geng, Jia
Subramaniam, Varuni
Lee, Tae Jin
Montemagno, Carlo
Guo, Peixuan
author_facet Wendell, David
Jing, Peng
Geng, Jia
Subramaniam, Varuni
Lee, Tae Jin
Montemagno, Carlo
Guo, Peixuan
author_sort Wendell, David
collection PubMed
description Biological pores have been used to study the transport of DNA and other molecules but most pores have channels that allow only the movement of small molecules and single-stranded DNA and RNA. The bacteriophage phi29 DNA-packaging motor, which allows double-stranded DNA to enter and exit during a viral infection, contains a connector protein that has a 3.6 – 6.0 nm wide channel. Here we show that a modified version of the connector protein, when reconstituted into liposomes and inserted into planar lipid bilayers, can act as conductive channels to allow the translocation of double-stranded DNA. Single-channel conductance assays and quantitative PCR confirmed the translocation through the pore. The measured conductance of a single connector channel was 4.8 nS in 1 M KCl. This engineered and membrane-adapted phage connector is expected to have interesting applications in nanotechnology and nanomedicine, such as MEMS sensing, microreactors, gene delivery, drug loading, and DNA sequencing.
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spelling pubmed-27777432010-05-01 Translocation of double stranded DNA through membrane adapted phi29 motor protein nanopore Wendell, David Jing, Peng Geng, Jia Subramaniam, Varuni Lee, Tae Jin Montemagno, Carlo Guo, Peixuan Nat Nanotechnol Article Biological pores have been used to study the transport of DNA and other molecules but most pores have channels that allow only the movement of small molecules and single-stranded DNA and RNA. The bacteriophage phi29 DNA-packaging motor, which allows double-stranded DNA to enter and exit during a viral infection, contains a connector protein that has a 3.6 – 6.0 nm wide channel. Here we show that a modified version of the connector protein, when reconstituted into liposomes and inserted into planar lipid bilayers, can act as conductive channels to allow the translocation of double-stranded DNA. Single-channel conductance assays and quantitative PCR confirmed the translocation through the pore. The measured conductance of a single connector channel was 4.8 nS in 1 M KCl. This engineered and membrane-adapted phage connector is expected to have interesting applications in nanotechnology and nanomedicine, such as MEMS sensing, microreactors, gene delivery, drug loading, and DNA sequencing. 2009-09-27 2009-11 /pmc/articles/PMC2777743/ /pubmed/19893523 http://dx.doi.org/10.1038/nnano.2009.259 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Wendell, David
Jing, Peng
Geng, Jia
Subramaniam, Varuni
Lee, Tae Jin
Montemagno, Carlo
Guo, Peixuan
Translocation of double stranded DNA through membrane adapted phi29 motor protein nanopore
title Translocation of double stranded DNA through membrane adapted phi29 motor protein nanopore
title_full Translocation of double stranded DNA through membrane adapted phi29 motor protein nanopore
title_fullStr Translocation of double stranded DNA through membrane adapted phi29 motor protein nanopore
title_full_unstemmed Translocation of double stranded DNA through membrane adapted phi29 motor protein nanopore
title_short Translocation of double stranded DNA through membrane adapted phi29 motor protein nanopore
title_sort translocation of double stranded dna through membrane adapted phi29 motor protein nanopore
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2777743/
https://www.ncbi.nlm.nih.gov/pubmed/19893523
http://dx.doi.org/10.1038/nnano.2009.259
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