School of BioSciences, The University of Melbourne, Parkville, VIC, Australia.
Methods Mol Biol. 2024;2775:81-90. doi: 10.1007/978-1-0716-3722-7_6.
Transformation of foreign DNA into Cryptococcus species is a powerful tool for exploring gene functions in these human pathogens. Agrobacterium tumefaciens-mediated transformation (AtMT) has been used for the stable introduction of exogenous DNA into Cryptococcus for over two decades, being particularly impactful for insertional mutagenesis screens to discover new genes involved in fungal biology. A detailed protocol to conduct this transformation method is provided in the chapter. Scope for modifications and the benefits and disadvantages of using AtMT in Cryptococcus species are also presented.
将外源 DNA 转化为隐球菌是探索这些人类病原体中基因功能的有力工具。农杆菌介导的转化(AtMT)已被用于将外源 DNA 稳定地导入隐球菌超过二十年,对于发现参与真菌生物学的新基因的插入突变筛选尤其有影响。本章提供了进行这种转化方法的详细方案。还介绍了在隐球菌中使用 AtMT 的修改范围以及优缺点。