Development of Impala-Based Transposon Systems for Gene Tagging in Filamentous Fungi

Abstract

Genome sequences of many filamentous fungi are now available and additional genomes are currently being sequenced. One of the next strategic goals is to generate collections of tagged genes in order to establish a link between the several thousands of predicted genes and their function. Transposable elements have been invaluable for the identification and isolation of genes of interest as insertion of a transposon both disrupts and tags a gene. In an effort to exploit active transposons identified in the genome of Fusarium oxysporum as insertional mutagens, a binary system including the tagging element, MITE, and the transposase of a Tc1 element has been established as an efficient tool for gene-tagging in Fusarium graminearum. In this chapter, we provide an overview of the techniques used and highlight some of the critical steps for the application of this tool to other fungal species.

Minos as a novel Tc1/mariner-type transposable element for functional genomic analysis in Aspergillus nidulans

“…To detect whether Minos transposes in A. nidulans we followed a strategy previously employed for the impala element (Li Destri Nicosia et al, 2001;Dufresne and Daboussi, 2010). The general idea is to introduce, by homologous recombination, a modified Minos element hosting a direct selection marker, in the promoter of the niaD gene, encoding nitrate reductase.…”

Advances in Molecular Tools and In Vivo Models for the Study of Human Fungal Pathogenesis

“…Transposon mutagenesis is also used in gene tagging thus facilitating systems-level analyses of non-model organisms [126][127][128][129]. In a recent study, a universal collection of Gateway-compatible 4280 TagModules was constructed as an efficient tool to generate tagged mutants in a range of microbial species.…”

“…Genetic studies in non-model fungi including various fungal pathogens are limited as the number of techniques available for generation and identification of gene mutations are limited. To overcome this, a number of autonomous and non-autonomous transposable elements have used to develop genetic tools to study fungal pathogens [ 128 , 129 , 131 , 132 ]. For example, an engineered Aspergillus fumigatus strain containing Impala160 , a DNA transposon from Fusarium oxysporum was employed to generate and screen a library of insertional mutants.…”

Γενετική τροποποίηση του μύκητα Fusarium Oxysporum με στόχο τη δημιουργία στελεχών με αυξημένη παραγωγικότητα αιθανόλης

“…Τα μεταθετά στοιχεία μπορούν να χρησιμοποιηθούν στη δημιουργία βιβλιοθηκών, με στόχο τη συσχέτιση των γονιδίων με τη λειτουργία τους, καθώς η εισαγωγή τους μέσα σε κάποιο γονίδιο διακόπτει τη λειτουργία του και ταυτόχρονα το επισημαίνει. Αυτή η δυνατότητα έχει ήδη διερευνηθεί στο F. graminearum και μπορεί να εφαρμοστεί και σε άλλα είδη μυκήτων (Dufresne and Daboussi, 2010).…”