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The RNA-dependent RNA polymerase, QDE-1, is a component of the RNA silencing pathway in Neurospora crassa. The enzymatically active carboxy-terminal fragment QDE-1 DeltaN has been expressed in Saccharomyces cerevisiae in the presence and absence of selenomethionine (SeMet). The level of SeMet incorporation was estimated by mass spectrometry to be approximately 98%. Both native and SeMet proteins were crystallized in space group P2(1) with unit cell parameters a=101.2, b=122.5, c=114.4A, beta=108.9 degrees , and 2 molecules per asymmetric unit. The native and SeMet crystals diffract to 2.3 and 3.2A, respectively, the latter are suitable for MAD structure determination.

Original publication

DOI

10.1016/j.jsb.2004.10.001

Type

Journal article

Journal

J Struct Biol

Publication Date

01/2005

Volume

149

Pages

111 - 115

Keywords

Cloning, Molecular, Crystallization, Crystallography, X-Ray, Fungal Proteins, Gene Silencing, Neurospora crassa, Peptide Fragments, RNA Replicase, Saccharomyces cerevisiae, Selenomethionine