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. |