ArticleFungal biology2013
Spectrum of T-DNA integrations for insertional mutagenesis of Histoplasma capsulatum.
Article in Fungal biology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
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- Tools for genetic manipulation of the endemic fungal pathogen,bioRxiv : the preprint server for biology · 2025Article
- Genotypic diversity, virulence, and molecular genetic tools inMicrobiology and molecular biology reviews : MMBR · 2024Review
- Targeted gene deletions in the dimorphic fungal pathogenmSphere · 2023Article
- Optimizing Transformation Frequency ofJournal of fungi (Basel, Switzerland) · 2021Review
- Updates inJournal of fungi (Basel, Switzerland) · 2021Review
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- CaNRT2.1 Is Required for Nitrate but Not Nitrite Uptake in Chili Pepper PathogenFrontiers in microbiology · 2020Article
- Generation ofMicroorganisms · 2019Article
- Macrophage activation by IFN-γ triggers restriction of phagosomal copper from intracellular pathogens.PLoS pathogens · 2018Article
- The use of3 Biotech · 2018Article
- O-Mannosylation of Proteins EnablesmBio · 2018Article
- Agrobacterium tumefaciens-mediated transformation: an efficient tool for targeted gene disruption in Talaromyces marneffei.World journal of microbiology & biotechnology · 2017Article
- Agrobacterium-mediated insertional mutagenesis in the mycorrhizal fungus Laccaria bicolor.Current genetics · 2017Article
- Eng1 and Exg8 Are the Major β-Glucanases Secreted by the Fungal PathogenThe Journal of biological chemistry · 2017Article
- Low-affinity iron transport protein Uvt3277 is important for pathogenesis in the rice false smut fungus Ustilaginoidea virens.Current genetics · 2017Article
- A silver bullet in a golden age of functional genomics: the impact ofFungal biology and biotechnology · 2017Review
- Revisiting old friends: Developments in understanding Histoplasma capsulatum pathogenesis.Journal of microbiology (Seoul, Korea) · 2016Review
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3 authors.
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Abstract
Agrobacterium-mediated transformation is being increasingly used for insertional mutagenesis of fungi. To better evaluate its effectiveness as a mutagen for the fungal pathogen Histoplasma capsulatum, we analyzed a collection of randomly selected T-DNA insertion mutants. Testing of different T-DNA element vectors engineered for transformation of fungi showed that pBHt2 provides the highest transformation efficiency and the lowest rate of vector backbone carryover. Sixty-eight individual T-DNA integrations were characterized by recovery of T-DNA ends and flanking genomic sequences. The right border (RB) end of the T-DNA is largely preserved whereas the left border (LB) end is frequently truncated. Analysis of T-DNA insertion sites confirms the lack of any integration hotspots in the Histoplasma genome. Relative to genes, T-DNA integrations show significant bias towards promoter regions at the expense of coding sequences. With consideration for potential promoter interruption and the demonstrated efficacy of intronic insertions, 61 % of mapped T-DNA insertions should impair gene expression or function. Mapping of T-DNA flanking sequences demonstrates 67 % of T-DNA integrations are integrations at a single chromosomal site and 31 % of T-DNA integrations are associated with large-scale chromosomal rearrangements. This characterization of T-DNA insertions in mutants selected without regard to phenotype supports application of Agrobacterium-mediated transformation as an insertional mutagen for genome-based screens and functional discovery of genes in Histoplasma.
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