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首页> 外文期刊>Archives of microbiology >A site-directed integration system for the nonuniversal CUG(Ser) codon usage species Pichia farinosa by electroporation
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A site-directed integration system for the nonuniversal CUG(Ser) codon usage species Pichia farinosa by electroporation

机译：通过电穿孔的非通用CUG（Ser）密码子使用种粉菌毕赤酵母的定点整合系统

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摘要

Halotolerant yeast, Pichia farinosa, is a valuable yeast strain in fermentation industry because it produces high yield of glycerol and xylitol, and can tolerate both contamination and high-density growth during fermentation. However, the lack of genetic manipulation tools makes it less popular as a gene engineering strain. Expression systems commonly used in other yeast systems, such as Saccharomyces cerevisiae and Pichia pastoris cannot be used in P. farinosa because it translates universal Leu codon CUG as Ser. Here we reported a modified expression vector and a transformation system with enhanced efficiency in P. farinosa. The results showed that cells of OD600 0.8-1.0 with DTT treatment can obtain high transformation efficiency. The optimized electroporation condition was 900 V, 25 mu F, and 200 Omega. The DNA concentration did not influence the transformation. Our system provides the potential not only for applying P. farinosa as an industrial strain of gene engineering, but also for studying gene function in its native host.
机译：耐盐酵母毕赤酵母（Pichia farinosa）是发酵工业中有价值的酵母菌株，因为它可产生高产量的甘油和木糖醇，并且可以耐受发酵过程中的污染和高密度生长。但是，由于缺乏基因操作工具，因此它不适合作为基因工程菌株。在其他酵母系统中常用的表达系统，例如酿酒酵母和巴斯德毕赤酵母，不能用于粉状假单胞菌，因为它会将通用Leu密码子CUG转换为Ser。在这里，我们报道了一种修饰的表达载体和一种转化系统，具有较高的效率。结果表明，经DTT处理的OD600为0.8-1.0的细胞可获得较高的转化效率。优化的电穿孔条件为900 V，25μF和200Ω。 DNA浓度不影响转化。我们的系统不仅为将粉状疟原虫用作基因工程的工业菌株提供了潜力，而且为研究其天然宿主中的基因功能提供了潜力。

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