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      Utilizing Yellow Fluorescent Protein Tagging to Determine Location of Histone H3.1 Protein in Arabidopsis thaliana

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            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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            Author and article information

            Journal
            ScienceOpen Posters
            ScienceOpen
            21 September 2022
            Affiliations
            [1 ] Biological Sciences, California State University, Chico, CA, USA
            Author notes
            Author information
            https://orcid.org/0000-0002-3017-1981
            Article
            10.14293/S2199-1006.1.SOR-.PPCQ7TB.v1
            f23caff8-bf8a-4bdf-82b4-062dbdbdf9fe

            This work has been published open access under Creative Commons Attribution License CC BY 4.0 , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Conditions, terms of use and publishing policy can be found at www.scienceopen.com .

            History
            : 21 September 2022

            All data generated or analysed during this study are included in this published article (and its supplementary information files).
            General life sciences,Plant science & Botany,Cell biology,Ecology,Molecular biology,Genetics
            tri-template PCR,PCR,epigenetics,subcellular localization,cell biology,molecular biology,protein localization,fluorescent protein,nuclear localization signal,histone protein

            References

            1. Hruz Tomas, Laule Oliver, Szabo Gabor, Wessendorp Frans, Bleuler Stefan, Oertle Lukas, Widmayer Peter, Gruissem Wilhelm, Zimmermann Philip. Genevestigator V3: A Reference Expression Database for the Meta-Analysis of Transcriptomes. Advances in Bioinformatics. Vol. 2008:1–5. 2008. Hindawi Limited. [Cross Ref]

            2. Nakai Kenta, Horton Paul. PSORT: a program for detecting sorting signals in proteins and predicting their subcellular localization. Trends in Biochemical Sciences. Vol. 24(1):34–35. 1999. Elsevier BV. [Cross Ref]

            3. Berardini Tanya Z., Reiser Leonore, Li Donghui, Mezheritsky Yarik, Muller Robert, Strait Emily, Huala Eva. The arabidopsis information resource: Making and mining the “gold standard” annotated reference plant genome. genesis. Vol. 53(8):474–485. 2015. Wiley. [Cross Ref]

            4. Talbert Paul B., Henikoff Steven. Histone variants on the move: substrates for chromatin dynamics. Nature Reviews Molecular Cell Biology. Vol. 18(2):115–126. 2017. Springer Science and Business Media LLC. [Cross Ref]

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