ArticleMolecular and biochemical parasitology
A densely overlapping gene fragmentation approach improves yeast two-hybrid screens for Plasmodium falciparum proteins.
Article in Molecular and biochemical parasitology. 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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6 citing papers in PubMed.
- The Plasmodium falciparum MESA erythrocyte cytoskeleton-binding (MEC) motif binds to erythrocyte ankyrin.Molecular and biochemical parasitology · 2019Article
- Article
- The Plasmodium falciparum exported protein PF3D7_0402000 binds to erythrocyte ankyrin and band 4.1.Molecular and biochemical parasitology · 2017Article
- Intrinsic disorder mediates hepatitis C virus core-host cell protein interactions.Protein science : a publication of the Protein Society · 2015Article
- Identification and comparative analysis of hepatitis C virus-host cell protein interactions.Molecular bioSystems · 2013Article
- A physical interaction network of dengue virus and human proteins.Molecular & cellular proteomics : MCP · 2011Article
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Abstract
Use of the yeast two-hybrid assay to study Plasmodium falciparum protein-protein interactions is limited by poor expression of P. falciparum genes in yeast and lack of easily implemented assays to confirm the results. We report here two methods to create gene fragments - random fragmentation by partial DNAse I digestion and generation of densely overlapping fragments by PCR - that enable most portions of P. falciparum genes to be expressed and screened in the yeast two-hybrid assay. The PCR-based method is less technically challenging and facilitates fine-scale mapping of protein interaction domains. Both approaches revealed a putative interaction between PfMyb2 (PF10_0327) and PFC0365w. We developed new plasmids to express the proteins in wheat germ extracts and confirmed the interaction in both the split-luciferase assay and in co-purification experiments with glutathione-S-transferase and HA-tagged proteins. The combination of improved yeast two-hybrid screening approaches and convenient systems to validate interactions enhances the utility of yeast two-hybrid assays for P. falciparum.
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