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SSO1450 - A CAS1 protein from Sulfolobus solfataricus P2 with high affinity for RNA and DNA
Authors:Dong Han
Institution:Department of Biochemistry, Universität Bayreuth, Universitätsstrasse 30, Bayreuth 95447, Germany
Abstract:Clustered regularly interspaced short palindromic repeats (CRISPR) and their associated protein genes (cas genes) are ubiquitous in archaea and eubacteria. It has been suggested that CRISPR and CAS proteins act as an immune system preventing the invasion of foreign genomic elements at the DNA level. The protein SSO1450 from Sulfolobus solfataricus (Sso) P2 belongs to the CAS1 cluster which is one of the core protein clusters most frequently associated with CRISPR sequences. In this study we show that SSO1450 is a high-affinity nucleic acid binding protein. It binds DNA, RNA and DNA-RNA hybrid apparently sequence non-specific in a multi-site binding mode. Furthermore, SSO1450 promotes the hybridization of complementary nucleic acid strands.
Keywords:CAS  CRISPR-associated  CASS  CRISPR-associated system  COG  cluster of orthologous groups  CRISPR  clustered regularly interspaced short palindromic repeats  DTT  dithiothreitol  EMSA  electrophoretic mobility shift assay  IPTG  d-thiogalactoside" target="_blank">isopropyl-β-d-thiogalactoside  Kd  apparent dissociation constant  LB  Luria-Bertani medium  OD600  optical density at 600   nm  PAGE  polyacrylamide gel electrophoresis  RNAi  RNA interference  SDS  sodium dodecylsulfate  Sso  Sulfolobus solfataricus  ss/ds  single/double-stranded  bp  base pair  nt  nucleotide
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