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在Cordyceps militaris中使用CRISPR/Cas9核糖蛋白传递有效地破坏CmHk1

Hyeongju Choi¹, Sung-Won Park¹, Junsang Oh²

¹Department of Integrative Food, Bioscience and Biotechnology, Chonnam National University, Gwangju 61186, Korea.

FEMS microbiology letters

|July 21, 2023

概括

我们开发了一种CRISPR/Cas9基因编辑系统,用于药用Cordyceps militaris. 该系统成功修改了CmHk1基因,揭示了它在杀菌剂耐药性和透应激反应中的作用.

科学领域:

菌类学菌类学是指菌类学.
遗传学是一个遗传学.
生物技术是生物技术.

背景情况:

科尔迪塞普斯军人是一种有价值的虫病原菌,具有药用特性.
开发高效的基因操纵工具对于理解和改进C. militaris.至关重要.
以前用于C. militaris的遗传转换的方法是有限的.

研究的目的:

建立一个强大的CRISPR/Cas9核糖核蛋白 (RNP) 传递系统,用于Cordyceps militaris的遗传转化.
调查C. militaris.中的III类胺激酶基因 (CmHk1) 的功能.
分析CmHk1淘汰对真菌杀菌剂敏感性和透应激耐受性的影响.

主要方法:

绿色光蛋白 (GFP) 突变体的生成使用PEG介导的原始质细胞转化.
将针对CmHk1的CRISPR/Cas9 RNP传递到C. militaris原生体中.
突变的特征和结果突变的体外灵敏度测试.

主要成果:

成功开发和实施CRISPR/Cas9 RNP交付系统.
识别CmHk1中的各种突变,包括删除和插入.
在CmHk1淘汰突变中,对杀菌剂iprodione和fludioxonil的敏感性降低.
与野生类型相比,CmHk1淘汰突变体对透应激的敏感性增加.

结论:

开发的CRISPR/Cas9 RNP系统对Cordyceps militaris的基因工程有效.
在C. militaris中,CmHk1基因在调解杀剂耐药性和对透应激反应方面发挥着重要作用.
这项研究为进一步对这种重要的药用进行功能基因组学研究提供了基础.

关键词:

科尔迪塞普斯军用菌 (Cordyceps militaris) 是一种植物.这就是CRISPR/Cas9的作用.迪卡博西米德真菌杀菌剂希斯提丁激酶的作用是核蛋白是一种核蛋白.

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