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康宁木霉原生质体的制备及质粒DNA转化方案。

Protocol for generating protoplasts of Trichoderma koningiopsis and transformation of plasmid DNA.

作者信息

Jin Sophie, Alberti Fabrizio

机构信息

School of Life Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, UK.

School of Life Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, UK.

出版信息

STAR Protoc. 2025 Mar 21;6(1):103668. doi: 10.1016/j.xpro.2025.103668. Epub 2025 Mar 6.

DOI:10.1016/j.xpro.2025.103668
PMID:40053449
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11930090/
Abstract

Trichoderma koningiopsis is a filamentous fungus that produces numerous bioactive molecules and has the potential to be used as a biological control and plant growth-promoting agent. Here, we present a protocol for generating protoplasts of T. koningiopsis. We then describe four techniques for the stable transformation of plasmid DNA: polyethylene glycol (PEG)-mediated transformation of protoplasts, Agrobacterium-mediated transformation, electroporation of spores, and lipofection of mycelia. We then detail procedures for confirming the expression of a spCas9 gene using fluorescence microscopy.

摘要

康宁木霉是一种丝状真菌，能产生多种生物活性分子，有潜力用作生物防治和植物生长促进剂。在此，我们展示了一种制备康宁木霉原生质体的方法。然后我们描述了四种用于质粒DNA稳定转化的技术：聚乙二醇（PEG）介导的原生质体转化、农杆菌介导的转化、孢子电穿孔和菌丝体脂质体转染。接着我们详细说明了使用荧光显微镜确认spCas9基因表达的程序。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/c9e97ccb608e/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/602a1a24c8f7/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/530608ddff10/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/b8881dbc3f22/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/8b093bb9b240/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/c9e97ccb608e/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/602a1a24c8f7/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/530608ddff10/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/b8881dbc3f22/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/8b093bb9b240/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d007/11930090/c9e97ccb608e/gr4.jpg

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