Biological Function of RMR2 in Maize: Genetic Study through Fluorescent Tagging of RMR2 Protein

Li, Han
Recent research in maize genomics shows that heritable chromatin changes contribute to an epigenetic process called gene silencing. A protein called RMR2 (required to maintain repression 2) is useful to study the process because mutations in the gene that encodes RMR2 prevent reversion of the epigenetic change. Phenotypic results of RMR2 mutants suggest the protein may silence maize pigmentation production through transcriptional gene silencing (TGS). This project aims to study the function of the RMR2 protein by localizing its expression during development and in relation to other proteins involved in TGS. The Gateway method for gene tagging is being used to produce a visual marker line. The method uses two cloning recombination reactions: First, BP reactions produce donor vectors for each of the selected segments of the genomic regions of the rmr2 gene, including regulatory regions and the sequence for the fluorescent probe (citrine YFP). LR reactions then recombine the plasmid donor vectors to make the final construct. After sequencing confirmation, the construct is transformed into a bacterial vector for maize transformation. RMR2 protein function and localization will then be studied using a laser scanning confocal microscope. We hypothesize that RMR2 interacts in a protein complex, including RMR1, to facilitate chromatin remodeling.
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