隐球酵母vps41Δ菌株转化条件优化及突变体库构建

Optimization of Agrobacterium-mediated Transformation of Cryptococcus neoformans vps41Δ and Construction of a Insertional mutagenesis Library

新型隐球酵母(Cryptococcus neoformans)是一种重要的条件致病性真菌。前期试验证实VPS41基因参与隐球酵母的氮源饥饿应答反应并影响其致病性。系统研究了影响农杆菌介导的vps41Δ饥饿应答缺陷型菌株关键因素,并利用优化的农杆菌转化体系构建了包含5 000多个转化子的vps41Δ菌株的T-DNA插入突变体库,为下一步筛选鉴定隐球酵母VPS41饥饿应答信号通路中的其他相关基因提供了基础材料。

Cryptococcus neoformans is a major human opportunistic fungal pathogens. The previous work has shown that VPS41 gene contributes to virulence by mediating nitrogen starvation response. In this study, major factors affecting the efficiency of Agrobacterium tumefaciens-mediated transformation of vps41Δ were systematically investigated, and optimized parameters were used to construct an insertional metagenesis library of the deletion mutant containing over 5 000 transformants. The availability of this insertional library could facilite the screening and identification of other genes involved in the signaling pathway of starvation response of C. neoformans.