RRC ID 40089
著者 Toh-E A, Ohkusu M, Shimizu K, Kawamoto S.
タイトル Positional cloning in Cryptococcus neoformans and its application for identification and cloning of the gene encoding methylenetetrahydrofolate reductase.
ジャーナル Fungal Genet Biol
Abstract Cryptococcus neoformans, a basidiomycetous human pathogenic yeast, has been widely used in research fields in medical mycology as well as basic biology. Gene cloning or identification of the gene responsible for a mutation of interest is a key step for functional analysis of a particular gene. The availability therefore, of the multiple methods for cloning is desirable. In this study, we proposed a method for a mapping-based gene identification/cloning (positional cloning) method in C. neoformans. To this end, we constructed a series of tester strains, one of whose chromosomes was labeled with the URA5 gene. A heterozygous diploid constructed by crossing one of the tester strains to a mutant strain of interest loses a chromosome(s) spontaneously, which is the basis for assigning a recessive mutant gene to a particular chromosome in the mitotic mapping method. Once the gene of interest is mapped to one of the 14 chromosomes, classical genetic crosses can then be performed to determine its more precise location. The positional information thus obtained can then be used to significantly narrow down candidate genes by referring to the Cryptococcus genome database. Each candidate gene is then examined whether it would complement the mutation. We successfully applied this method to identify CNA07390 encoding methylenetetrahydrofolate reductase as the gene responsible for a methionine-requiring mutant in our mutant collection.
巻・号 76
ページ 70-7
公開日 2015-3-1
DOI 10.1016/j.fgb.2015.02.007
PII S1087-1845(15)00022-5
PMID 25687932
MeSH Cloning, Molecular / methods Cryptococcus neoformans / enzymology Cryptococcus neoformans / genetics* Methylenetetrahydrofolate Reductase (NADPH2) / chemistry Methylenetetrahydrofolate Reductase (NADPH2) / genetics* Mutation
IF 3.071
引用数 3
WOS 分野 MYCOLOGY GENETICS & HEREDITY
リソース情報
病原微生物 NA