A novel complex: a quantum dot conjugated to an active T7 RNA Polymerase

Mette Eriksen; Peter Horvath; Michael Askvad Sørensen; Szabolcs Semsey; Lene Broeng Oddershede; Liselotte Jauffred Pedersen

doi:10.1155/2013/468105

A novel complex: a quantum dot conjugated to an active T7 RNA Polymerase

Mette Eriksen, Peter Horvath, Michael Askvad Sørensen, Szabolcs Semsey, Lene Broeng Oddershede, Liselotte Jauffred Pedersen

2 Citationer (Scopus)

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Abstract

To perform single-molecule studies of the T7 RNA polymerase, it is crucial to visualize an individual T7 RNA polymerase, for example, through a fluorescent signal. We present a novel complex combining two different molecular functions, an active T7 RNA polymerase and a highly luminescent nanoparticle, a quantum dot. The complex has the advantage of both constituents: the complex can traffic along DNA and simultaneously be visualized, both at the ensemble and at the single-molecule level. The labeling was mediated through an in vivo biotinylation of a His-tagged T7 RNA polymerase and subsequent binding of a streptavidin-coated quantum dot. Our technique allows for easy purification of the quantum dot labeled T7 RNA polymerases from the reactants. Also, the conjugation does not alter the functionality of the polymerase; it retains the ability to bind and transcribe.

Originalsprog	Engelsk
Artikelnummer	468105
Tidsskrift	Journal of Nanomaterials
Vol/bind	2013
Antal sider	9
ISSN	1687-4110
DOI	https://doi.org/10.1155/2013/468105
Status	Udgivet - 2013

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10.1155/2013/468105

A Novel Complex: A Quantum Dot Conjugated to an Active T7 RNA PolymeraseForlagets udgivne version, 3,49 MB

http://dx.doi.org/10.1155/2013/468105

Citationsformater

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abstract = "To perform single-molecule studies of the T7 RNA polymerase, it is crucial to visualize an individual T7 RNA polymerase, for example, through a fluorescent signal. We present a novel complex combining two different molecular functions, an active T7 RNA polymerase and a highly luminescent nanoparticle, a quantum dot. The complex has the advantage of both constituents: the complex can traffic along DNA and simultaneously be visualized, both at the ensemble and at the single-molecule level. The labeling was mediated through an in vivo biotinylation of a His-tagged T7 RNA polymerase and subsequent binding of a streptavidin-coated quantum dot. Our technique allows for easy purification of the quantum dot labeled T7 RNA polymerases from the reactants. Also, the conjugation does not alter the functionality of the polymerase; it retains the ability to bind and transcribe.",

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T2 - a quantum dot conjugated to an active T7 RNA Polymerase

AU - Eriksen, Mette

AU - Horvath, Peter

AU - Sørensen, Michael Askvad

AU - Semsey, Szabolcs

AU - Oddershede, Lene Broeng

AU - Pedersen, Liselotte Jauffred

PY - 2013

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N2 - To perform single-molecule studies of the T7 RNA polymerase, it is crucial to visualize an individual T7 RNA polymerase, for example, through a fluorescent signal. We present a novel complex combining two different molecular functions, an active T7 RNA polymerase and a highly luminescent nanoparticle, a quantum dot. The complex has the advantage of both constituents: the complex can traffic along DNA and simultaneously be visualized, both at the ensemble and at the single-molecule level. The labeling was mediated through an in vivo biotinylation of a His-tagged T7 RNA polymerase and subsequent binding of a streptavidin-coated quantum dot. Our technique allows for easy purification of the quantum dot labeled T7 RNA polymerases from the reactants. Also, the conjugation does not alter the functionality of the polymerase; it retains the ability to bind and transcribe.

AB - To perform single-molecule studies of the T7 RNA polymerase, it is crucial to visualize an individual T7 RNA polymerase, for example, through a fluorescent signal. We present a novel complex combining two different molecular functions, an active T7 RNA polymerase and a highly luminescent nanoparticle, a quantum dot. The complex has the advantage of both constituents: the complex can traffic along DNA and simultaneously be visualized, both at the ensemble and at the single-molecule level. The labeling was mediated through an in vivo biotinylation of a His-tagged T7 RNA polymerase and subsequent binding of a streptavidin-coated quantum dot. Our technique allows for easy purification of the quantum dot labeled T7 RNA polymerases from the reactants. Also, the conjugation does not alter the functionality of the polymerase; it retains the ability to bind and transcribe.

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DO - 10.1155/2013/468105

M3 - Journal article

SN - 1687-4110

VL - 2013

JO - Journal of Nanomaterials

JF - Journal of Nanomaterials

M1 - 468105

ER -

A novel complex: a quantum dot conjugated to an active T7 RNA Polymerase

Abstract

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