Genetic Manipulation of Cryptococcus neoformans.
Genetic Manipulation of Cryptococcus neoformans.
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DOI：
10.1002/cpmc.59
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发表时间：
2018-08-01
影响因子：
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通讯作者：
Bahn, Yong-Sun
中科院分区：
文献类型：
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作者：
Jung, Kwang-Woo;Lee, Kyung-Tae;Bahn, Yong-Sun
新型隐球菌是一种机会性真菌病原体，其在免疫功能低下的个体中引起危及生命的脑膜脑炎，并且每年在全世界造成超过1，000，000例感染和600，000例死亡。然而，抗隐球菌的治疗选择是有限的，主要是因为真菌和人类细胞结构之间的相似性。由于近十年来遗传、分子生物学技术和生物信息学的发展，C.属于担子菌门的neoformans是目前主要的致病真菌模式系统。特别是，遗传操作是识别和表征基因功能以理解致病性的基础机制的第一步。新人类本单元描述了使用双接头（DJ）PCR构建靶基因缺失突变体，使用组蛋白H3基因启动子构建组成型过表达菌株，以及在C.新人类John Wiley & Sons，Inc.
Cryptococcus neoformans is an opportunistic fungal pathogen, which causes life-threatening meningoencephalitis in immunocompromised individuals and is responsible for more than 1,000,000 infections and 600,000 deaths annually worldwide. Nevertheless, anti-cryptococcal therapeutic options are limited, mainly because of the similarity between fungal and human cellular structures. Owing to advances in genetic and molecular techniques and bioinformatics in the past decade, C. neoformans, belonging to the phylum basidiomycota, is now a major pathogenic fungal model system. In particular, genetic manipulation is the first step in the identification and characterization of the function of genes for understanding the mechanisms underlying the pathogenicity of C. neoformans. This unit describes protocols for constructing target gene deletion mutants using double-joint (DJ) PCR, constitutive overexpression strains using the histone H3 gene promoter, and epitope/fluorescence protein-tagged strains in C. neoformans. © 2018 by John Wiley & Sons, Inc.