title: Expression and Purification of the CMR (Type III-B) Complex in Sulfolobus solfataricus.
abstract: Protein purification is an important technique that allows us to characterize the structural and biochemical properties of either an individual protein or a multi-protein complex. However, expression and purification of one subunit of a complex in the absence of its binding partners has often proven difficult to achieve due to the issues such as instability and mis-folding. This is the case for the components of the CRISPR-Cas interference complexes, which degrade invading nucleic acids in a sequence homology-dependent manner in many prokaryotic species. Here, we describe the expression of a tandem-tagged subunit of the Type III-B (CMR) complex in Sulfolobus solfataricus and subsequent isolation and purification of the whole complex by affinity purification of the tagged subunit.
authors.affiliation: Biomedical Sciences Research Complex, University of St Andrews, North Haugh, St Andrews, KY16 9ST, UK, jz21@st-andrews.ac.uk.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Malcolm F
authors.initials: MF
authors.lastname: White
mesh_terms: D019843:Archaeal Proteins / Q000235:genetics* / Q000302:isolation & purification*; D003001:Cloning, Molecular; D018274:Electroporation; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D048229:Sulfolobus solfataricus / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D019843:Archaeal Proteins