ArticleBMC microbiology2009
Reverse genetics through random mutagenesis in Histoplasma capsulatum.
Article in BMC microbiology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
12 citing papers in PubMed, 16 citations in OpenAlex.
- Targeted gene deletions in the dimorphic fungal pathogenmSphere · 2023Article
- Article
- A silver bullet in a golden age of functional genomics: the impact ofFungal biology and biotechnology · 2017Review
- Rapid mapping of insertional mutations to probe cell wall regulation in Cryptococcus neoformans.Fungal genetics and biology : FG & B · 2015Article
- Thermally Dimorphic Human Fungal Pathogens--Polyphyletic Pathogens with a Convergent Pathogenicity Trait.Cold Spring Harbor perspectives in medicine · 2014Review
- The spectrum of fungi that infects humans.Cold Spring Harbor perspectives in medicine · 2014Review
- Glycosylation and immunoreactivity of the Histoplasma capsulatum Cfp4 yeast-phase exoantigen.Infection and immunity · 2014Article
- Quantitative analysis of deadenylation-independent mRNA decay by a modified MBRACE assay.Methods in molecular biology (Clifton, N.J.) · 2014Article
- Spectrum of T-DNA integrations for insertional mutagenesis of Histoplasma capsulatum.Fungal biology · 2013Article
- Histoplasma mechanisms of pathogenesis--one portfolio doesn't fit all.FEMS microbiology letters · 2011Review
- Discovery of a role for Hsp82 in Histoplasma virulence through a quantitative screen for macrophage lethality.Infection and immunity · 2011Article
- The yeast-phase virulence requirement for α-glucan synthase differs among Histoplasma capsulatum chemotypes.Eukaryotic cell · 2011Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
backgroundThe dimorphic fungal pathogen Histoplasma capsulatum causes respiratory and systemic disease in humans and other mammals. Progress in understanding the mechanisms underlying the biology and the pathogenesis of Histoplasma has been hindered by a shortage of methodologies for mutating a gene of interest.
resultsWe describe a reverse genetics process that combines the random mutagenesis of Agrobacterium-mediated transformation with screening techniques to identify targeted gene disruptions in a collection of insertion mutants. Isolation of the desired mutant is accomplished by arraying individual clones from a pool and employing a PCR-addressing method. Application of this procedure facilitated the isolation of a cbp1 mutant in a North American type 2 strain, a Histoplasma strain recalcitrant to gene knock-outs through homologous recombination. Optimization of cryopreservation conditions allows pools of mutants to be banked for later analysis and recovery of targeted mutants.
conclusionThis methodology improves our ability to isolate mutants in targeted genes, thereby facilitating the molecular genetic analysis of Histoplasma biology. The procedures described are widely applicable to many fungal systems and will be of particular interest to those for which homologous recombination techniques are inefficient or do not currently exist.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.