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Characterization of the RNA-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis,

R Soc Open Sci. 2021-04; 
Benjamin H Jenkins, Finlay Maguire, Guy Leonard, Joshua D Eaton, Steven West, Benjamin E Housden, David S Milner, Thomas A Richards
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PCR Cloning and Subcloning … Sequences for plasmid constructs were synthesized de novo by either Genscript or SynBio Technologies, and cloned into an L4440 plasmid vector (Addgene plasmid #1654). Sequences and cloning sites for each plasmid construct are detailed in electronic supplementary … Get A Quote

摘要

Endosymbiosis was fundamental for the evolution of eukaryotic complexity. Endosymbiotic interactions can be dissected through forward- and reverse-genetic experiments, such as RNA-interference (RNAi). However, distinguishing small (s)RNA pathways in a eukaryote-eukaryote endosymbiotic interaction is challenging. Here, we investigate the repertoire of RNAi pathway protein-encoding genes in the model nascent endosymbiotic system, spp. Using comparative genomics and transcriptomics supported by phylogenetics, we identify essential proteome components of the small interfering (si)RNA, scan (scn)RNA and internal eliminated sequence (ies)RNA pathways. Our analyses reveal that copies of these components have been ret... More

关键词

RNAi, algae, dicer, interaction, siRNA, symbiosis