AN OPTIMIZED METHOD FOR THE ISOLATION OF NUCLEI FROM CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE)(1)
© 2011 Phycological Society of America.
| Veröffentlicht in: | Journal of phycology. - 1966. - 47(2011), 2 vom: 29. Apr., Seite 333-40 |
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| Weitere Verfasser: | , , |
| Format: | Online-Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2011
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| Zugriff auf das übergeordnete Werk: | Journal of phycology |
| Schlagworte: | Journal Article 2D gel electrophoresis Chlamydomonas algae nuclear proteins nucleus proteomics |
| Zusammenfassung: | © 2011 Phycological Society of America. 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 |
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| Beschreibung: | Date Completed 30.03.2016 Date Revised 29.03.2016 published: Print-Electronic Citation Status PubMed-not-MEDLINE |
| ISSN: | 1529-8817 |
| DOI: | 10.1111/j.1529-8817.2011.00967.x |