ArticleProceedings of the National Academy of Sciences of the United States of America1992
Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1992. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 178 papers.
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Who cites it
178 citing papers in PubMed.
- Many Cells Make Light Work: Self-Generated Gradients OrganizeCold Spring Harbor perspectives in biology · 2026Review
- Unlocking CRISPR-Cas9 editing for widely diverse Dictyostelid species.Molecular systems biology · 2026Article
- A nanobody-based degron system for targeted protein knockdown in Dictyostelium discoideum.BMC molecular and cell biology · 2026Article
- Advantages of Mutant Generation by Genome Rearrangements of Non-Conventional Yeast via Direct Nuclease Transfection.Genes to cells : devoted to molecular & cellular mechanisms · 2025Article
- Mutation Screening of Dictyostelium Restriction Enzyme-Mediated Integration (REMI) Libraries.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Parallel evolution of the G protein-coupled receptor GrlG and the loss of fruiting body formation in the social amoeba Dictyostelium discoideum evolved under low relatedness.G3 (Bethesda, Md.) · 2023Article
- A phosphatidylinositol phosphate kinase inhibits Ras activation and regulates chemorepulsion in Dictyostelium discoideum.Journal of cell science · 2023Article
- Katnip is needed to maintain microtubule function and lysosomal delivery to autophagosomes and phagosomes.Molecular biology of the cell · 2023Article
- CRISPR/Cas9-based genome-wide screening of Dictyostelium.Scientific reports · 2022Article
- Engineering the amoeba Dictyostelium discoideum for biosynthesis of a cannabinoid precursor and other polyketides.Nature biotechnology · 2022Article
- Model-based identification of conditionally-essential genes from transposon-insertion sequencing data.PLoS computational biology · 2022Article
- Review
- The Amoebal Model for Macropinocytosis.Sub-cellular biochemistry · 2022Article
- Identification of novel proteins in themicroPublication biology · 2022Article
- A chemorepellent inhibits local Ras activation to inhibit pseudopod formation to bias cell movement away from the chemorepellent.Molecular biology of the cell · 2022Article
- Mutant resources for functional genomics in Dictyostelium discoideum using REMI-seq technology.BMC biology · 2021Article
- Review
- Development of a Positive Selection High Throughput Genetic Screen inFrontiers in cell and developmental biology · 2021Article
- Loss of PIKfyve Causes Transdifferentiation ofFrontiers in cell and developmental biology · 2021Article
- Article
118 more citing papers are in PubMed but not listed here.
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2 authors.
Funding
Abstract
Introduction of restriction enzyme along with linearized plasmid results in integration of plasmid DNA at genomic restriction sites in a high proportion of the resulting transformants. We have found that electroporating BamHI or EcoRI together with pyr5-6 plasmids cut with the same enzyme stimulates the efficiency of transformation in Dictyostelium discoideum more than 20-fold over the rate seen when plasmid DNA alone is introduced. Restriction enzyme-mediated integration generates insertions into genomic restriction sites in an apparently random manner, some of which cause mutations. About 1 in 400 of the Dictyostelium transformants displayed arrested or aberrant development. The integrated plasmid, along with flanking genomic DNA, was excised from some of these mutants, cloned in Escherichia coli, and used to transform other Dictyostelium cells. Homologous recombination within the flanking sequences resulted in the same phenotypes displayed by the original mutants, directly demonstrating that the affected genes were responsible for the specific morphological phenotypes. This method of insertional mutagenesis should be useful for tagging, and subsequent cloning, of many developmentally important genes that can be identified by their mutant phenotypes.
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