碩士 / 國立臺灣大學 / 微生物與生化學研究所 / 98 / SUMO (small ubiquitin-like modifier) is one of the ubiquitin-like proteins, which can covalently attach to target proteins for post-translational modification. SUMOylation cascade includes SUMO E1 activating enzyme, E2 conjugating enzyme and E3 ligases. SUMO E1 activating enzyme is a heterodimer composed of SAE1 (sumo activating enzyme subunit 1) and SAE2 (sumo activating enzyme subunit 2).
I initially performed the BLAST search by using the SUMO E1 sequences of Saccharomyces cerevisiae, Arabidopsis thaliana and human as the template, and predicted the potential exon-intron regions by AUGUSTUS program to find out the potential E1 activating enzyme of Chlamydomonas reinhardtii. In this study, the SAE1 and SAE2 partial sequences were successfully cloned by using specific primers respectively. The results showed that GST (glutathione S-transferase) can help the protein expression of SAE1 and SAE2 in Escherichia coli. The in vitro pull down assay did not reveal an interaction between SAE2 partial sequence and CrSUMO96-GG. Furthermore, I also used the GST-CrSAE2 partial sequence as the antigen for preparing the polyclonal antibody. Hopefully, it can be the specific tool for study of C. reinhardtii SUMO E1 activating enzyme in the future.
| Identifer | oai:union.ndltd.org:TW/098NTU05381146 |
| Date | January 2010 |
| Creators | Li-Hsien Huang, 黃俐嫻 |
| Contributors | 張世宗 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 64 |
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