AN OPTIMIZED METHOD FOR THE ISOLATION OF NUCLEI FROM CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE)

Flavia Vischi Winck, Miroslaw Kwasniewski, Stefanie Wienkoop (Corresponding author), Bernd Müller-Röber

    Publications: Contribution to journalArticlePeer Reviewed

    Abstract

    The cell nucleus harbors a large number of proteins involved in transcription, RNA processing, chromatin remodeling, nuclear signaling, and ribosome assembly. The nuclear genome of the model alga Chlamydomonas reinhardtii P. A. Dang. was recently sequenced, and many genes encoding nuclear proteins, including transcription factors and transcription regulators, have been identified through computational discovery tools. However, elucidating the specific biological roles of nuclear proteins will require support from biochemical and proteomics data. Cellular preparations with enriched nuclei are important to assist in such analyses. Here, we describe a simple protocol for the isolation of nuclei from Chlamydomonas, based on a commercially available kit. The modifications done in the original protocol mainly include alterations of the differential centrifugation parameters and detergent-based cell lysis. The nuclei-enriched fractions obtained with the optimized protocol show low contamination with mitochondrial and plastid proteins. The protocol can be concluded within only 3 h, and the proteins extracted can be used for gel-based and non-gel-based proteomic approaches.
    Original languageEnglish
    Pages (from-to)333-340
    Number of pages8
    JournalJournal of Phycology: an international journal of algal research
    Volume47
    Issue number2
    Publication statusPublished - 2011

    Austrian Fields of Science 2012

    • 106002 Biochemistry
    • 106037 Proteomics

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