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Nutrient supplements boost yeast transformation efficiency

机译:营养补充剂可提高酵母转化效率

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摘要

Efficiency of yeast transformation is determined by the rate of yeast endocytosis. The aim of this study was to investigate the effect of introducing amino acids and other nutrients (inositol, adenine, or p-aminobenzoic acid) in the transformation medium to develop a highly efficient yeast transformation protocol. The target of rapamycin complex 1 (TORC1) kinase signalling complex influences the rate of yeast endocytosis. TORC signaling is induced by amino acids in the media. Here, we found that increasing the concentration of amino acids and other nutrients in the growth media lead to an increase yeast transformation efficiency up to 107 CFU per μg plasmid DNA and per 108 cells with a 13.8 kb plasmid DNA. This is over 130 times that of current published methods. This improvement may facilitate more efficient experimentation in which transformation efficiency is critical, such as yeast two-hybrid screening.
机译:酵母转化的效率由酵母内吞率决定。这项研究的目的是研究在转化培养基中引入氨基酸和其他营养物质(肌醇,腺嘌呤或对氨基苯甲酸)对开发高效酵母转化方案的影响。雷帕霉素复合物1(TORC1)激酶信号转导复合物的目标影响酵母内吞率。 TORC信号传导是由培养基中的氨基酸诱导的。在这里,我们发现,增加生长培养基中氨基酸和其他营养素的浓度会导致酵母转化效率提高,每μg质粒DNA和每108个细胞中带有13.8 kb质粒的酵母转化效率最高可达107 CFU。这是当前已发布方法的130倍以上。这种改进可以促进更高效的实验,在该实验中,转化效率至关重要,例如酵母双杂交筛选。

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