Skip to content

Research at St Andrews

Expression and purification of the CMR (Type III-B) complex in Sulfolobus solfataricus

Research output: Contribution to journalArticlepeer-review

Author(s)

Jing Zhang, Malcolm F White

School/Research organisations

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.
Close

Details

Original languageEnglish
Pages (from-to)185-94
Number of pages10
JournalMethods in Molecular Biology
Volume1311
DOIs
Publication statusPublished - 2015

    Research areas

  • Affinity purification, Hyperthermophile, Tandem tags, Protein complex, Viral shuttle vector

Discover related content
Find related publications, people, projects and more using interactive charts.

View graph of relations

Related by author

  1. The CRISPR ancillary effector Can2 is a dual-specificity nuclease potentiating type III CRISPR defence

    Zhu, W., McQuarrie, S. J., Gruschow, S., McMahon, S., Graham, S., Gloster, T. & White, M., 15 Feb 2021, In: Nucleic Acids Research. Advance Article, 13 p., gkab073.

    Research output: Contribution to journalArticlepeer-review

  2. Facile and scalable expression and purification of transcription factor IIH (TFIIH) core complex

    Sanles-Falagan, R., Petrovic-Stojanovska, B. & White, M. F., Oct 2020, In: Protein Expression and Purification. 174, 105660.

    Research output: Contribution to journalArticlepeer-review

  3. Tetramerisation of the CRISPR ring nuclease Crn3/Csx3 facilitates cyclic oligoadenylate cleavage

    Athukoralage, J. S., McQuarrie, S. J., Gruschow, S., Graham, S., Gloster, T. & White, M., 20 Jul 2020, In: eLife. 9, 19 p., e57627.

    Research output: Contribution to journalArticlepeer-review

  4. Bacteria SAVED from viruses

    White, M. F., 9 Jul 2020, In: Cell. 182, 1, p. 5-6 2 p.

    Research output: Contribution to journalArticlepeer-review

  5. Fuse to defuse: a self-limiting ribonuclease-ring nuclease fusion for type III CRISPR defence

    Samolygo, A., Athukoralage, J. S., Graham, S. & White, M., 19 Jun 2020, In: Nucleic Acids Research. 48, 11, p. 6149–6156 8 p.

    Research output: Contribution to journalArticlepeer-review

Related by journal

  1. Functional studies of DNA-protein interactions using FRET techniques

    Blouin, S., Craggs, T. D., Lafontaine, D. A. & Penedo, C., 1 Oct 2015, In: Methods in Molecular Biology. 1334, p. 115-141

    Research output: Contribution to journalArticlepeer-review

  2. Molecular genetic methods to study DNA replication protein function in Haloferax volcanii, a model archaeal organism.

    Giroux, X. & MacNeill, S., 2015, In: Methods in Molecular Biology. 1300, p. 187-218 31 p.

    Research output: Contribution to journalArticlepeer-review

  3. Pumilio-based RNA in vivo imaging

    Tilsner, J., 2015, In: Methods in Molecular Biology. 1217, p. 295-328

    Research output: Contribution to journalArticlepeer-review

  4. Use of microarray analysis to investigate EMT gene signatures

    Sims, A. H., Larionov, A. A., Harrison, D. J. & Katz, E., 2013, In: Methods in Molecular Biology. 1046, p. 85-95 11 p.

    Research output: Contribution to journalArticlepeer-review

ID: 208699104

Top