使用酸/和载体DNA进行化学能力高的Pichia pastoris细胞的高吞吐量转化
Kirill Smirnov1, Amina Hasanagic2, Lukas Rieder3
1Christian Doppler Laboratory for Innovative Pichia pastoris Host and Vector Systems, Graz University of Technology, Petersgasse, Graz, Austria. k.smirnov@tugraz.at.
Methods in molecular biology (Clifton, N.J.)
|October 1, 2025
概括
一个使用酸/的新化学转化协议用于Komagataella phaffii提供了电穿孔的替代方案. 这种方法适用于当高转换效率不是关键时的并行处理.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 电穿孔是Komagataella phaffii转化的常见方法,提供高效率但有限的可扩展性.
- 96井格式对于使用电穿孔的并行转换是不方便的.
研究的目的:
- 描述生产具有化学能力的K. phaffii细胞的协议.
- 详细说明K. phaffii.基于酸/化物的转化协议.
- 提供适合特定应用的替代转化方法.
主要方法:
- 使用酸/化物生产化学上合格的K. phaffii细胞.
- 使用载体DNA与化学方法的K. phaffii的转化.
主要成果:
- 酸/化物方法提供了1 x 10^2和1 x 10^3 CFU/μg DNA之间的转换效率.
- 这种化学转换协议可以适应并行处理,与电穿孔不同.
结论:
- 酸/化物协议是K. phaffii转换的可行替代电穿孔方法.
- 这种方法在高转换率不是必不可少的情况下是有利的,例如在图书馆生成中.
- 该协议为96井格式的并行处理提供了更好的适应性.
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