Title [PROTOCOL]A Signature-Tagged Mutagenesis (STM)-based murine-infectivity assay for Cryptococcus neoformans
Author Kwang-Woo Jung1, Kyung-Tae Lee2, and Yong-Sun Bahn2*
Address 1Radiation Research Division, Korea Atomic Energy Research Institute, Jeongeup 56212, Republic of Korea, 2Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea
Bibliography Journal of Microbiology, 58(10),823–831, 2020,
DOI 10.1007/s12275-020-0341-8
Key Words fungal pathogens, quantitative PCR, virulence, mutant library
Abstract Signature-tagged mutagenesis (STM) is a high-throughput genetic technique that can be used to investigate the function of genes by constructing a large number of mutant strains with unique DNA identification tags, pooling them, and screening them for a particular phenotypic trait. STM was first designed for the identification of genes that contribute to the virulence or infectivity of a pathogen in its host. Recently, this method has also been applied for the identification of mutants with specific phenotypes, such as antifungal drug resistance and proliferation. In the present study, we describe an STM method for the identification of genes contributing to the infectivity of Cryptococcus neoformans using a mutant library, in which each strain was tagged with a unique DNA sequence.