Genetic Manipulation of Cryptococcusneoformans

摘要

Cryptococcus neoformans is an opportunistic fungal pathogen, which causeslife-threatening meningoencephalitis in immunocompromised individuals andis responsible for more than 1,000,000 infections and 600,000 deaths annuallyworldwide. 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 inthe past decade, C. neoformans, belonging to the phylum basidiomycota, isnow a major pathogenic fungal model system. In particular, genetic manipulationis the first step in the identification and characterization of the functionof genes for understanding the mechanisms underlying the pathogenicity ofC. neoformans. This unit describes protocols for constructing target gene deletionmutants using double-joint (DJ) PCR, constitutive overexpression strainsusing the histone H3 gene promoter, and epitope/fluorescence protein-taggedstrains in C. neoformans.