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Identification of Essential Genes in the Human Fungal Pathogen Aspergillus fumigatus by Transposon Mutagenesis

机译:转座子诱变鉴定人类真菌病原烟曲霉中的必需基因

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摘要

The opportunistic pathogen Aspergillus fumigatus is the most frequent cause of deadly airborne fungal infections in developed countries. In order to identify novel antifungal-drug targets, we investigated the genome of A. fumigatus for genes that are necessary for efficient fungal growth. An artificial A. fumigatus diploid strain with one copy of an engineered impala160 transposon from Fusarium oxysporum integrated into its genome was used to generate a library of diploid strains by random in vivo transposon mutagenesis. Among 2,386 heterozygous diploid strains screened by parasexual genetics, 1.2% had a copy of the transposable element integrated into a locus essential for A. fumigatus growth. Comparison of genomic sequences flanking impala160 in these mutants with that of the genome of A. fumigatus allowed the characterization of 20 previously uncharacterized A. fumigatus genes. Among these, homologues of genes essential for Saccharomyces cerevisiae growth have been identified, as well as genes that do not have homologues in other fungal species. These results confirm that heterologous transposition using the transposable element impala is a powerful tool for functional genomics in ascomycota, and they pave the way for defining the complete set of essential genes in A. fumigatus, the first step toward target-based development of new antifungal drugs.
机译:在发达国家,机会病原体烟曲霉是致命的空气传播真菌感染的最常见原因。为了鉴定新型抗真菌药物靶标,我们调查了烟曲霉的基因组,寻找有效真菌生长所需的基因。人工烟曲霉二倍体菌株具有一个拷贝的来自尖孢镰刀菌(Fusarium oxysporum)的工程Impala160转座子的拷贝整合到其基因组中,用于通过随机体内转座子诱变产生二倍体菌株的文库。在2,386个通过双性恋遗传学筛选的杂合二倍体菌株中,有1.2%的转座因子拷贝整合到了烟曲霉生长必不可少的基因座中。将这些突变体中的impala160侧翼的基因组序列与烟曲霉的基因组序列进行比较,可以表征20个以前未表征的烟曲霉基因。其中,已鉴定出酿酒酵母生长所必需的基因的同源物,以及在其他真菌物种中不具有同源物的基因。这些结果证实,使用转座子Impala进行异源转座是子囊菌功能基因组学的强大工具,它们为定义烟曲霉完整的必需基因集铺平了道路,这是朝目标靶点开发新型抗真菌剂的第一步毒品。

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