Isolation and analysis of Arabidopsis thaliana lines bearing DRG knock-out mutations and overexpressing GFP protein fusions: Are wild-type DRGs necessary for heat stress granule formation?
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Purpose and Methodology: The purpose of this study was to determine the role of DRG and GFP in response to heat stress in Arabidopsis by examining whether any DRG mutant combination inhibits the formation of heat stress granules (HSGs) following exposure of plants to heat stress. Methods utilized in the experimentation include the polymerase chain reaction (PCR) and gel electrophoresis to screen for plants containing the desired combination of genes, a root-growth assay to study the relationship between phenotype and genetic composition, and confocal microscopy to observe the formation of HSGs in root tips. Findings: The genetic composition of DRG genes and GFPs does have an effect on the phenotype of Arabidopsis. Conversely, the presence of a wild type DRG gene fused to GFP (DRGI-GFP or DRG2-GFP) does not complement the non-functional DRG genes.