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Utilizing Yellow Fluorescent Protein Tagging to Determine Location of Histone H3.1 Protein in Arabidopsis thaliana
Abstract
The objective of this study was to identify the final destination of the Histone H3.1 protein in the root tip of Arabidopsis thaliana plants. Histones are present in heterochromatin in the form of nucleosomes. Consequently, identifying H3 protein location provides insight into its cellular interactions and functions in the root tip of A. thaliana. H3.1 protein subcellular localization will be accomplished via yellow fluorescent protein (YFP) tagging technique. I hypothesize that the insertion of YFP between the core and tail of the histone gene will result in a fluorescent H3.1 protein located in the nucleus of a transgenic plant. Tri-Template PCR was used to incorporate YFP into the At1g09200 gene (an H3.1 gene). The plant embryo was inoculated with Agrobacterium containing the linearized H3.1-YFP transgene. The transgene would translate into a protein with a fluorescent tag. Areas of fluorescence during plant growth could be recorded to identify the location of the protein within the root of the transgenic plant. Observations of plant growth could not be recorded due to circumstances. Therefore, genetic databases were referenced to formulate predictions of where the H3.1 protein will fluoresce in the plant cell and plant root. We predict fluorescence in the root apical meristem of the plant and at the periphery of the nucleus, indicating the presence of H3.1 protein in heterochromatin in the form of nucleosomes. Localization of the H3.1 protein using YFP tagging will facilitate our exploration of histone protein functions such as DNA packaging and transcriptional regulation.
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References
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