The Pseudozyma flocculosa actin promoter allows the strong expression of a recombinant protein in the Pseudozyma species
The Pseudozyma flocculosa actin promoter allows the strong expression of a recombinant protein in the Pseudozyma species
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DOI：
10.1007/s00253-006-0786-7
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发表时间：
2007-04-01
影响因子：
5
通讯作者：
Belanger, Richard R.
中科院分区：
文献类型：
--
作者：
Neveu, Bertrand;Michaud, Melanie;Belanger, Richard R.
属于新近归类的假酵母属的真菌具有独特的生物活性、产生稀有的抗菌糖脂和产生重组蛋白等特性。在这项工作中，我们首次克隆了肌动蛋白基因的内源启动子，该启动子是多面酵母样絮状假酵母的内源启动子。启动子区域缺乏典型的TATA或CAAT盒，但有三个推测的GC盒和两个CT富集区。与其他相关担子菌一样，丛枝假单胞菌基因组中只有一个肌动蛋白基因拷贝。比较了肌动蛋白启动子和玉米黑粉菌HSP70启动子在两种拟酵母菌中的活性。与液体培养中HSP70启动子相比，肌动蛋白启动子在丛生红曲霉中可以表达非常高的GFP蛋白(27.8mgg(-1)总蛋白)。相比之下，在液体培养中获得的绿色荧光蛋白的表达水平与南极假酵母菌的肌动蛋白或HSP70启动子相似。在固体培养中也观察到类似的GFP表达模式。这一新启动子的克隆为进一步开发和研究假酵母菌属真菌的特殊性质提供了独特的遗传工具。
Fungi belonging to the recently classified genus Pseudozyma possess some unique properties such as biocontrol activity, production of rare antimicrobial glycolipids and production of recombinant proteins. In this work, we report the first cloning of a promoter endogenous to the multi-faceted yeast-like Pseudozyma flocculosa, that of the actin gene. The promoter region lacked typical TATA or CAAT box but displayed three putative GC box and two CT-rich regions. As in other related basidiomycetes, only one copy of the actin gene was present in the genome of P flocculosa. The activity of the actin promoter was compared to that of the HSP70 promoter from Ustilago maydis in two Pseudozyma species. In R flocculosa, the actin promoter allowed the expression of a very high amount of GFP protein (27.8 mg g(-1) total protein) compared to those obtained with the HSP70 promoter in liquid culture. By contrast, the levels of GFP expression obtained in liquid culture were similar with the actin or the HSP70 promoter in Pseudozyma antarctica. A similar pattern of GFP expression was observed in solid culture. The cloning of this new promoter offers a unique genetic tool to further exploit and study the unusual properties of fungi from the Pseudozyma genus.