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Type III CRISPR complexes from Thermus thermophilus

Szychowska, Marta and Siwek, Wojciech and Pawolski, Damian and Kazrani, Asgar Abbas and Pyrc, Krzysztof and Bochtler, Matthias (2016) Type III CRISPR complexes from Thermus thermophilus. Acta Biochimica Polonica, 63 (2). pp. 377-386. ISSN 1734-154X

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Abstract

Pathogen-specific acquired immunity in bacteria is mediated by the CRISPR (clustered regularly interspaced short palindromic repeats)-Cas systems. Thermus thermophilus strain HB8 contains CRISPR systems of several major subtypes (type I, IIIA and IIIB), and has become a widely studied model for CRISPR biology. We have selected two highly expressed CRISPR spacers, crRNA 2.1 and crRNA 2.2, and have enriched endogenous T. thermophilus proteins that co-purify with these crRNAs. Mass spectroscopy indicates that the chromatography protocol enriches predominantly Csm complex subunits, but also Cmr subunits. After several chromatographic steps, size exclusion chromatography indicated a molecular mass of the crRNA associated complex of 265±69 kDa. In agreement with earlier work, crRNAs of different lengths (containing the selected spacers) were observed. Most of these were completely lost when several T. thermophilus csm genes were ablated.

Item Type:Article
Subjects:Q Science > Q Science (General)
Divisions:Department of Bioinformatics
ID Code:1273
Deposited By: Prof. Matthias Bochtler
Deposited On:23 Dec 2016 10:37
Last Modified:23 Dec 2016 10:37

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