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 Citations (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.

Original language	English
Article number	468105
Journal	Journal of Nanomaterials
Volume	2013
Number of pages	9
ISSN	1687-4110
DOIs	https://doi.org/10.1155/2013/468105
Publication status	Published - 2013

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A Novel Complex: A Quantum Dot Conjugated to an Active T7 RNA PolymeraseFinal published version, 3.49 MB

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

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title = "A novel complex: a quantum dot conjugated to an active T7 RNA Polymerase",

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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doi = "10.1155/2013/468105",

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volume = "2013",

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T1 - A novel complex

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

Y1 - 2013

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.

U2 - 10.1155/2013/468105

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

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