| 崔铁忠 卢康荣 李银霞 王万山.酿酒酵母遗传缺陷型菌株化学转化方法研究[J].,2016,16(8):1420-1423 |
| 酿酒酵母遗传缺陷型菌株化学转化方法研究 |
| Research on a Chemical-Based Yeast Transformation Method for DefectiveSaccharomyces Cerevisiae |
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| DOI： |
| 中文关键词: 酵母 遗传转化 醋酸锂 |
| 英文关键词: Yeast Transformation Lithiumacetate |
| 基金项目:广东省科技计划项目（2011B060300028; 2012B040304010） |
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| 中文摘要: |
| 目的：通过改善转化条件，提高缺陷型酿酒酵母DNA转化效率。方法：以醋酸锂化学转化法为基础，以酿酒酵母菌株::
KAN作为遗传转化受体，以敲除基因为转化目的，对影响遗传缺陷型菌株转化效率的参数（醋酸锂前处理、热激时间及转
化后复苏时间）进行优化，确定适合缺陷型酵母DNA 转化的最佳方法。结果：不同时间（30 min，60 min 和120 min）醋酸锂的前处
理均可以提高其转化效率，热激前使用醋酸锂处理酵母30 min 转化效率最高；热激10 min 是转化效率由高到低的转折点，30 min
热激明显降低了转化效率；YPD培养基用于转化后酵母的复苏，随培养时间（30 min，60 min 和120 min）的延长，酵母转化效率逐
步增加，120 min 作为热敏性菌株转化后的复苏时间较佳。结论：与野生型菌株类似，通过转化条件的优化，遗传缺陷型菌株的转
化效率也可以满足大多数试验要求。 |
| 英文摘要: |
| Objective:To improve the transformation efficiency of defective yeast strains by testing different transformation
conditions.Methods:Based on LiAc-transformation method, strain ::KAN was used as genetic
transformation receptor, and the transformation was done by knocking out gene, then the best conditions for transforming
defective yeast strains were determined by optimizing the transformation parameters such as LiAc pretreatment, heat shock time and
post-transformation recovery time.Results:Different time (30 min, 60 min and 120 min following LiAc treatment) could all improve the
transformation efficiency, of which the highest efficiency can be achieved by 30 min pretreatment. 10 min after heat shock was the
turning point from high transformation efficiency to low ones, and 30 min heat shock reduced efficiency remarkably. Transformation
efficiency gradually increased along with the prolongation of culture time (30 min, 60 min and 120 min) when YPD medium was used to
recover the post-transformation yeast, 120 min recovery in YPD medium should be preferred after heat shock.Conclusion:By
improving transformation conditions, the transformation of defective yeast strains can satisfy the need of most purposes. |
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