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  Enzymatic ligation strategies for the preparation of purine riboswitches with site-specific chemical modifications.

Rieder, R., Hoebartner, C., & Micura, R. (2009). Enzymatic ligation strategies for the preparation of purine riboswitches with site-specific chemical modifications. In A. Serganov (Ed.), Riboswitches: Methods and protocols (pp. 15-24). New York, N.Y.: Humana Pr. doi:10.1007/978-1-59745-558-9_2.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0013-0EE3-1 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-1430-3
Genre: Book Chapter

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603127.pdf (Publisher version), 323KB
 
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 Creators:
Rieder, R., Author
Hoebartner, C.1, Author              
Micura, R., Author
Affiliations:
1Research Group of Nucleic Acid Chemistry, MPI for biophysical chemistry, Max Planck Society, ou_578605              

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Free keywords: Ligation - Riboswitch - Aminopurine - RNA modification
 Abstract: One of the most versatile riboswitch classes refers to purine nucleoside metabolism. In the cell, purine riboswitches of the respective mRNAs either act at the transcriptional or translational level and off- or on-regulate genes upon binding to their dedicated ligands. Biophysical studies on ligand-induced folding of these RNA domains in vitro contribute to understanding their regulation mechanisms in vivo. For such studies, in particular, for approaches using fluorescence spectroscopy, the preparation of large RNAs with site-specific chemical modifications is required. Here, we describe a strategy for the preparation of riboswitch aptamers and aptamers adjoined to their expression platforms by chemical synthesis and enzymatic ligation. The modular design enables fast access to a large number of purine riboswitch derivatives with the modification of interest at any strand position. We exemplarily provide a detailed protocol for the preparation of adenosine deaminase (add) A-riboswitch variants with 2-aminopurine (AP) modifications at the 40-nmol scale.

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Language(s): eng - English
 Dates: 2009-05-012009
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: eDoc: 430538
DOI: 10.1007/978-1-59745-558-9_2
 Degree: -

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Source 1

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Title: Riboswitches: Methods and protocols
Source Genre: Book
 Creator(s):
Serganov, A., Editor
Affiliations:
-
Publ. Info: New York, N.Y. : Humana Pr.
Pages: XI, 366 Volume / Issue: - Sequence Number: - Start / End Page: 15 - 24 Identifier: ISBN: 978-1-934115-88-6

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Title: Methods in molecular biology
Source Genre: Series
 Creator(s):
Walker, J. M., Editor
Affiliations:
-
Publ. Info: -
Pages: - Volume / Issue: 540 Sequence Number: - Start / End Page: - Identifier: -