A codon-optimized luciferase from Gaussia princeps facilitates the in vivo monitoring of gene expression in the model alga Chlamydomonas reinhardtii

被引:86
作者
Shao, Ning [1 ]
Bock, Ralph [1 ]
机构
[1] Max Planck Inst Mol Pflanzenphysiol, D-14476 Potsdam, Germany
关键词
reporter gene; Chlamydomonas reinhardtii; luciferase; Gaussia princeps; heat-inducible expression; bioluminescence;
D O I
10.1007/s00294-008-0189-7
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The unicellular green alga Chlamydomonas reinhardtii has emerged as a superb model species in plant biology. Although the alga is easily transformable, the low efficiency of transgene expression from the Chlamydomonas nuclear genome has severely hampered functional genomics research. For example, poor transgene expression is held responsible for the lack of sensitive reporter genes to monitor gene expression in vivo, analyze subcellular protein localization or study protein-protein interactions. Here, we have tested the luciferase from the marine copepod Gaussia princeps (G-Luc) for its suitability as a sensitive bioluminescent reporter of gene expression in Chlamydomonas. We show that a Gaussia luciferase gene variant, engineered to match the codon usage in the Chlamydomonas nuclear genome, serves as a highly sensitive reporter of gene expression from both constitutive and inducible algal promoters. Its bioluminescence signal intensity greatly surpasses previously developed reporters for Chlamydomonas nuclear gene expression and reaches values high enough for utilizing the reporter as a tool to monitor responses to environmental stresses in vivo and to conduct high-throughput screenings for signaling mutants in Chlamydomonas.
引用
收藏
页码:381 / 388
页数:8
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