增强酵母菌转化：通过在醋酸锂-聚乙二醇方法中的一次调整实现高达十倍的增加。

IF 2.2 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Yeast Pub Date : 2025-03-15 DOI:10.1002/yea.3999

Mathilde Kadouch, Pierre Gaspin, Christelle Marchal, Sabine Castano, Christophe Cullin

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引用次数: 0

摘要

锂- peg转化酵母细胞的方法是大多数酵母实验室使用的标准程序。经过多次优化，该方法每µg质粒可产生多达106个转化子。一些应用程序，如库筛选或复杂的转换，需要最大化转换产量。在这里，我们证明添加山梨糖醇溶液在热冲击期间和之后作为渗透保护剂，导致转化效率提高10倍。与原始方案相比，这种优化只需要一个额外的移液步骤，使其适合日常使用。

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Enhancing Yeast Transformation: Achieving up to a Tenfold Increase Through a Single Adjustment in the Lithium Acetate-Polyethylene Glycol Method.

The Lithium-PEG method for transforming yeast cells is a standard procedure used in most yeast laboratories. After several optimizations, this method can yield up to 10⁶ transformants per µg of plasmid. Some applications, such as library screening or complex transformations, necessitate maximizing transformation yield. Here, we demonstrate that the addition of a sorbitol solution serves as an osmo-protectant during and after heat shock, resulting in up to a tenfold increase in transformation efficiency. This optimization requires only one additional pipetting step compared to the original protocol, making it practical for routine use.

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来源期刊

Yeast 生物-生化与分子生物学

CiteScore

5.30

自引率

3.80%

发文量

审稿时长

3 months

期刊介绍： Yeast publishes original articles and reviews on the most significant developments of research with unicellular fungi, including innovative methods of broad applicability. It is essential reading for those wishing to keep up to date with this rapidly moving field of yeast biology. Topics covered include: biochemistry and molecular biology; biodiversity and taxonomy; biotechnology; cell and developmental biology; ecology and evolution; genetics and genomics; metabolism and physiology; pathobiology; synthetic and systems biology; tools and resources