Kawakami, H., Stillman, B. (December 2012) In vitro reconstitution of budding yeast pre-RC, competent in recruitment of recombinant pre-IC components, using purified, recombinant proteins. Genes & Genetic Systems, 87 (6). p. 385. ISSN 1341-7568
Abstract
Pre-RC formation on chromosomal replication origins is an essential process for eukaryotic DNA replication in order for the origins to form pre-IC complexes. Though purification of each pre-RC component is of great importance to conduct quantitative analyses in vitro, recombinant MCM in budding yeast was a challenge to purify as a form active in pre-RC formation. We purified recombinant MCM complexed with Cdt1 and then reconstituted pre-RC formation using only purified, recombinant proteins. The established reaction could occur on representative early and late origins and was repressed by certain origin mutations. Reconstituted pre-RC had affinity for recombinant pre-IC components, indicating that a recombinant pre-IC could be assembled in vitro.
| Item Type: | Paper |
|---|---|
| Additional Information: | Meeting Abstract |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification Publication Type > Meeting Abstract bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > origin recognition complex bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types organism description > yeast |
| CSHL Authors: | |
| Communities: | CSHL labs > Stillman lab |
| Highlight: | Stillman, Bruce W. |
| Depositing User: | Matt Covey |
| Date: | December 2012 |
| Date Deposited: | 25 Sep 2013 15:36 |
| Last Modified: | 21 Feb 2018 20:49 |
| URI: | https://repository.cshl.edu/id/eprint/28623 |
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