Novel insights in genetic transformation of the probiotic yeast Saccharomyces boulardii
Novel insights in genetic transformation of the probiotic yeast Saccharomyces boulardii
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DOI：
10.4161/bioe.26271
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发表时间：
2014-01-01
期刊：
影响因子：
4.9
通讯作者：
Marques, Ernesto T. A., Jr.
中科院分区：
文献类型：
--
作者：
Douradinha, Bruno;Reis, Viviane C. B.;Marques, Ernesto T. A., Jr.
布拉酵母（Saccharomyces boulardii）（S.布拉氏酵母（S.boulardii）是与酿酒酵母（S.cerevisiae）相关的益生菌酵母。酿酒酵母），但具有不同的遗传、分类和代谢特性。S.酿酒酵母已广泛用于生物技术应用。目前，许多菌株是可用的，并且已经开发了多种遗传工具，其允许表达几种感兴趣的外源蛋白质，其应用于医学、生物燃料、食品工业和科学研究等领域。虽然S.布拉酵母由于其针对几种胃肠道疾病的益生菌特性而被广泛研究，但很少有研究涉及使用该酵母作为表达具有生物技术应用的感兴趣的外源基因的载体。在这里，我们表明，尽管两种酵母的相似性，并不是所有的遗传工具在S。酿酒酵母可应用于S.布拉地。虽然后者的转化可以使用为前者开发的商业试剂盒获得，但随后对成功转化体的筛选必须进行优化。我们还发现了几个在S.酿酒酵母（例如，启动子和抗性标记）存在于S.布拉地。测序显示这两种酵母的直系同源物之间有很高的同源性（>96%）。然而，我们也观察到其中一些不适合作为S.布拉地。这项工作对这种益生菌酵母作为感兴趣的基因的表达系统的潜力具有重要的应用。
Saccharomyces boulardii (S. boulardii) is a probiotic yeast related to Saccharomyces cerevisiae (S. cerevisiae) but with distinct genetic, taxonomic, and metabolic properties. S. cerevisiae has been used extensively in biotechnological applications. Currently, many strains are available, and multiple genetic tools have been developed, which allow the expression of several exogenous proteins of interest with applications in the fields of medicine, biofuels, the food industry, and scientific research, among others. Although S. boulardii has been widely studied due to its probiotic properties against several gastrointestinal tract disorders, very few studies addressed the use of this yeast as a vector for expression of foreign genes of interest with biotechnological applications. Here we show that, despite the similarity of the two yeasts, not all genetic tools used in S. cerevisiae can be applied in S. boulardii. While transformation of the latter could be obtained using a commercial kit developed for the former, consequent screening of successful transformants had to be optimized. We also show that several genes frequently used in genetic manipulation of S. cerevisiae (e.g., promoters and resistance markers) are present in S. boulardii. Sequencing revealed a high rate of homology (>96%) between the orthologs of the two yeasts. However, we also observed some of them are not eligible to be targeted for transformation of S. boulardii. This work has important applications toward the potential of this probiotic yeast as an expression system for genes of interest.