Characterization of the Protein Subcellular Localization and Promoter Specificity of OSCK1 from Rice
| Autor: | Pei-Ching Tsai, 蔡佩菁 |
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| Rok vydání: | 2002 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 90 The calcium-dependent calmodulin-independent protein kinases (CDPKs) are calcium-regulated protein kinases. The CDPKs are encoded by multigene family, although highly conserved in sequence, they often exhibit distinct expression patterns, implicating their specific roles involved in various developmental controls or physiological responses. By differential screening, we have isolated a CDPK gene, designated as OSCK1 (Oryza sativa calcium-dependent calmodulin-independent protein kinase 1), from rice. Database searching revealed that an myristoylation signal is located at the N-terminal end of OSCK1 and may be responsible for its membrane association. Moreover, a bipartite nucleus localization signal was found at the C-terminal of OSCK1. We aim to analyze the relevance of these two putative signals in directing the subcellular distribution and the biochemical activity of OSCK1. Our results showed that the OSCK1 was myristoylated when co-expressed with yeast N-myristoyl transferase (yNMT) in E. coli. However, no difference in kinase activity could be detected between proteins expressed with or without yNMT. Moreover, OSCK1-GUS fusion protein was not specifically accumulated in nucleus when transiently expressed in the epidermis of onion. To analyze the activity and specificity of OSCK1 promoter, we constructed three plasmids carrying OSCK1 promoter in various lengths fused with the GUS reporter gene. By transformation of rice, the preliminary results showed that no GUS activity could be detected in the vegetative tissues of transgenic plants. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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