A Novel Complex: A Quantum Dot Conjugated to an Active T7 RNA Polymerase. (19th May 2013)

• Record Type: Journal Article
• Authors: Eriksen, Mette; Horvath, Peter; Sørensen, Michael A.; Semsey, Szabolcs; Oddershede, Lene B.; Jauffred, Liselotte
• Other Names: Merlin Christophe Academic Editor.
• Is Part Of: Journal of nanomaterials. Volume 2013(2013)
• Publication Date: 2013-05-19
• Subjects: Nanostructured materials -- Periodicals; Nanotechnology -- Periodicals; Nanomatériaux; Nanostructured materials; Nanotechnology; Nanostructures; Nanotechnology; Periodicals; Fulltext; Internet Resources; Periodicals; 620.115
• DOI: 10.1155/2013/468105
• Languages: English
• ISSNs: 1687-4110

Abstract

Abstract : To perform single-molecule studies of the T 7 RNA polymerase, it is crucial to visualize an individual T 7 RNA polymerase, for example, through a fluorescent signal. We present a novel complex combining two different molecular functions, an active T 7 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 T 7 RNA polymerase and subsequent binding of a streptavidin-coated quantum dot. Our technique allows for easy purification of the quantum dot labeled T 7 RNA polymerases from the reactants. Also, the conjugation does not alter the functionality of the polymerase; it retains the ability to bind and transcribe.