Monitoring dynamic expression of nuclear genes in Chlamydomonas reinhardtii by using a synthetic luciferase reporter gene
| Autor: | Markus Fuhrmann, Amparo Hausherr, Lars Ferbitz, Thomas Schödl, Markus Heitzer, Peter Hegemann |
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| Rok vydání: | 2004 |
| Předmět: |
Time Factors
Recombinant Fusion Proteins Ribulose-Bisphosphate Carboxylase Algal Proteins Photosynthetic Reaction Center Complex Proteins Imidazoles Gene Expression Nuclear Proteins Plant Science General Medicine Transfection Substrate Specificity Pyrazines Enzyme Stability Genetics Escherichia coli Animals Luciferases Promoter Regions Genetic Agronomy and Crop Science Chlamydomonas reinhardtii Plasmids |
| Zdroj: | Plant molecular biology. 55(6) |
| ISSN: | 0167-4412 |
| Popis: | For monitoring the expression profile of selected nuclear genes in Chlamydomonas reinhardtii in response to altered environmental parameters or during cell cycle, in the past many RNA or protein samples had to be taken and analyzed by RNA hybridization or protein immunoblotting. Here we report the synthesis of a gene that codes for the luciferase of Renilla reniformis (RLuc) and is adapted to the nuclear codon usage of C. reinhardtii . This crluc gene was expressed alone or as a fusion to the zeocin resistance gene ble under control of different promoter variants. Luciferase activity was monitored in living cells, increased with the promoter strength and paralleled the amount of expressed protein. Under control of the Lhcb-1 promoter the Luc-activity in synchronized cultures was dependent on the dark-light cycle. Additionally, crluc was placed under control of the Chop-2 promoter and activity was measured under different light conditions. Chop-2 promoter activity was found to be most pronouced under low-light and dark conditions, further supporting that channelrhodopsin-2 is most active in dark-adapted cells. We conclude that crluc is a reliable tool for convenient monitoring of nuclear gene expression in C. reinhardtii . |
| Databáze: | OpenAIRE |
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