TY  - JOUR
A1  - Winck, Flavia Vischi
A1  - Kwasniewski, Miroslaw
A1  - Wienkoop, Stefanie
A1  - Müller-Röber, Bernd
T1  - An optimized method for the isolation of nuclei from chlamydomas Reinhardtii (Chlorophyceae)
JF  - Journal of phycology
N2  - 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.
KW  - 2D gel electrophoresis
KW  - algae
KW  - Chlamydomonas
KW  - nuclear proteins
KW  - nucleus
KW  - proteomics
Y1  - 2011
U6  - https://doi.org/10.1111/j.1529-8817.2011.00967.x
SN  - 0022-3646
VL  - 47
IS  - 2
SP  - 333
EP  - 340
PB  - Wiley-Blackwell
CY  - Malden
ER  -