Development of ToxA and ToxB promoter-driven fluorescent protein expression vectors for use in filamentous ascomycetes
Development of ToxA and ToxB promoter-driven fluorescent protein expression vectors for use in filamentous ascomycetes
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DOI：
10.3852/mycologia.97.5.1152
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发表时间：
2005-09-01
期刊：
影响因子：
2.8
通讯作者：
Ciuffetti, LM
中科院分区：
文献类型：
--
作者：
Andrie, RM;Martinez, JP;Ciuffetti, LM
绿色荧光蛋白（GFP）已被确立为研究基因表达、蛋白定位以及细胞和生物体动力学的首要体内报告者。真菌转化载体pCT 74，与sGFP的ToxA启动子的控制下，从Pyrenophora triticipentis，有效地表达GFP在一个不同的组的丝状子囊菌。由于ToxA启动子驱动的GFP表达的多功能性，我们构建了一组额外的荧光蛋白表达载体，以扩展用于丝状真菌的荧光标记物的调色板。EYFP、ECFP和mRFP 1从ToxA启动子成功地在其起源真菌P. tritici-repentis和远亲大丽轮枝菌（Verticilliumdahliae）中表达。此外，来自小麦疫霉的ToxB启动子驱动sGFP在大丽轮枝菌中的表达，表明ToxA启动子在子囊菌中异源表达的类似潜力。这里提出的真菌转化载体套件有望用于各种真菌研究应用。
The green fluorescent protein (GFP) has been established as the premier in vivo reporter for investigations of gene expression, protein localization, and cell and organism dynamics. The fungal transformation vector pCT74, with sGFP under the control of the ToxA promoter from Pyrenophora triticirepentis, effectively expresses GFP in a diverse group of filamentous ascomycetes. Due to the versatility of ToxA promoter-driven expression of GFP, we constructed an additional set of fluorescent protein expression vectors to expand the color palette of fluorescent markers for use in filamentous fungi. EYFP, ECFP and mRFP1 were successfully expressed from the ToxA promoter in its fungus of origin, P. tritici-repentis, and a distant relative, Verticillium dahliae. Additionally the ToxB promoter from P. tritici-repentis drove expression of sGFP in V. dahliae, suggesting a similar potential to the ToxA promoter for heterologous expression in ascomycetes. The Suite of fungal transformation vectors presented here promise to be useful for a variety of fungal research applications.