ArticleBMC microbiology2006
Optimization of a large-scale gene disruption protocol in Dictyostelium and analysis of conserved genes of unknown function.
Article in BMC microbiology, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- CRISPR/Cas9-based genome-wide screening of Dictyostelium.Scientific reports · 2022Article
- Review
- An RNA-binding protein, RNP-1, protects microtubules from nocodazole and localizes to the leading edge during cytokinesis and cell migration in Dictyostelium cells.Acta pharmacologica Sinica · 2016Article
- Article
- Dictyostelium discoideum--a model for many reasons.Molecular and cellular biochemistry · 2009Review
- Vacuole membrane protein 1 is an endoplasmic reticulum protein required for organelle biogenesis, protein secretion, and development.Molecular biology of the cell · 2008Article
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Authors and funding
3 authors.
Funding
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Abstract
backgroundDevelopment of the post-genomic age in Dictyostelium will require the existence of rapid and reliable methods to disrupt genes that would allow the analysis of entire gene families and perhaps the possibility to undertake the complete knock-out analysis of all the protein-coding genes present in Dictyostelium genome.
resultsHere we present an optimized protocol based on the previously described construction of gene disruption vectors by in vitro transposition. Our method allows a rapid selection of the construct by a simple PCR approach and subsequent sequencing. Disruption constructs were amplified by PCR and the products were directly transformed in Dictyostelium cells. The selection of homologous recombination events was also performed by PCR. We have constructed 41 disruption vectors to target genes of unknown function, highly conserved between Dictyostelium and human, but absent from the genomes of S. cerevisiae and S. pombe. 28 genes were successfully disrupted.
conclusionThis is the first step towards the understanding of the function of these conserved genes and exemplifies the easiness to undertake large-scale disruption analysis in Dictyostelium.
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