title: Targeted gene deletion in Aspergillus fumigatus using microbial machinery and a recyclable marker.
abstract: The emerging invasive fungal pathogen Aspergillus fumigatus causes very serious infections among immunocompromised patient populations. While the genome of this pathogen has been sequenced, a major barrier to better understanding the complex biology of this eukaryotic organism is a lack of tools for efficient genetic manipulation. To improve upon this, we have generated a new gene deletion system for A. fumigatus using yeast recombinational cloning and Agrobacterium tumefaciens mediated transformation (ATMT) employing a recyclable marker system. This system reduced the time for generating a gene deletion strain in our hands by two-thirds (12 weeks to 3 weeks) using minimal human labor, and we demonstrate that it can be used to efficiently generate multiple gene deletions within a single strain.
authors.affiliation: Department of Pathology, University of Iowa Carver College of Medicine, 200 Hawkins Drive, Iowa City, IA 52242, United States.
authors.forename: Joshua B
authors.initials: JB
authors.lastname: Kieler
authors.forename: Khanh L
authors.initials: KL
authors.lastname: Duong
authors.forename: W Scott
authors.initials: WS
authors.lastname: Moye-Rowley
authors.forename: J Stacey
authors.initials: JS
authors.lastname: Klutts
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D001232:Aspergillus fumigatus / Q000235:genetics*; D055786:Gene Knockout Techniques / Q000379:methods*; D005819:Genetic Markers; D005827:Genetics, Microbial / Q000379:methods*; D012641:Selection, Genetic; D013997:Time Factors; D015003:Yeasts / Q000235:genetics
chemical_list: D005819:Genetic Markers
keywords: Agrobacterium; Amylase; Aspergillus fumigatus; Gene deletion