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首页> 外文期刊>Journal of Biotechnology >Construction and characterization of centromeric, episomal and GFP-containing vectors for Saccharomyces cerevisiae prototrophic strains
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Construction and characterization of centromeric, episomal and GFP-containing vectors for Saccharomyces cerevisiae prototrophic strains

机译：酿酒酵母原养型菌株着丝粒，游离型和含GFP的载体的构建和表征

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摘要

A set of shuttle yeast vectors containing the dominant selectable markers KanMX4 or HphMX4 cassettes, conferring resistance to geneticin and hygromycin B, respectively, was constructed. Dominant selectable markers are useful for genetic manipulation of natural, wine and industrial strains which do not contain any auxotrophic markers as well as of strains which cannot grow on synthetic mineral medium. Vectors were characterized by (i) copy number, (ii) mitotic stability both in selective and non-selective conditions, (iii) the efficiency and frequency of transformations, (iv) optimal adaptation times in non-selective media, (v) optimum conditions for transformation of various laboratory, commercial and wine strains, and (vi) expression level of an inserted gene. Furthermore we produced GFP-containing vectors that can be used for protein subcellular localization in prototrophic strains.
机译：构建了一组穿梭酵母载体，其包含分别赋予遗传霉素和潮霉素B抗性的显性选择标记KanMX4或HphMX4盒。显性选择标记可用于不包含任何营养缺陷标记的天然，葡萄酒和工业菌株以及不能在合成矿物培养基上生长的菌株的遗传操作。载体的特征在于（i）拷贝数，（ii）在选择性和非选择性条件下的有丝分裂稳定性，（iii）转化的效率和频率，（iv）在非选择性培养基中的最佳适应时间，（v）最佳各种实验室，商业和葡萄酒菌株的转化条件，以及（vi）插入基因的表达水平。此外，我们生产了可用于原养菌株中蛋白质亚细胞定位的含GFP载体。

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