Present address: University of Vienna, Department of Molecular Systems Biology, Althanstraße 14, 1090 Vienna, Austria.
AN OPTIMIZED METHOD FOR THE ISOLATION OF NUCLEI FROM CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE)1
Article first published online: 21 MAR 2011
© 2011 Phycological Society of America
Journal of Phycology
Volume 47, Issue 2, pages 333–340, April 2011
How to Cite
Winck, F. V., Kwasniewski, M., Wienkoop, S. and Mueller-Roeber, B. (2011), AN OPTIMIZED METHOD FOR THE ISOLATION OF NUCLEI FROM CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE). Journal of Phycology, 47: 333–340. doi: 10.1111/j.1529-8817.2011.00967.x
Received 17 October 2009. Accepted 13 September 2010.
- Issue published online: 4 APR 2011
- Article first published online: 21 MAR 2011
- 2D gel electrophoresis;
- nuclear proteins;
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.