Cytoplasmic and Nuclear Localization of DcHSP17.7, a Small Heat Shock Protein
2007
Ahn, Y.J. (Sangmyung University, Seoul, Republic of Korea), E-mail: [email protected] | Zimmerman, J. Lynn (University of Maryland, Baltimore County, Baltimore, MD, USA)
The cellular localization of DcHSP17.7, a small heat shock protein (smHSP) from carrot (Daucus carota L.), was studied using green fluorescent protein (GFP) as a marker. A fusion gene construct, the DcHSP17.7 gene fused with the GFP gene, driven by the cauliflower mosaic virus 35S promoter was introduced into potato (Solanum tuberosum L. cv. 'Desiree') via Agrobacterium-mediated transformation. Transgenic plants were successfully regenerated and the DcHSP17.7-GFP fusion protein was observed using confocal laser scanning microscopy. Our results showed that DcHSP17.7 is localized in the cytoplasm and nucleus of the cells in leaf and stem tissues. Its general cellular compartmentation did not alter after heat stress (40℃, 4 hours). However, DcHSP17.7 was present at a higher level in the perinuclear region of the cytoplasm than inside the nucleus in the heat-stressed cells of stem tissues. The possible roles of DcHSP17.7 in enhancement of thermotolerance under heat stress conditions were discussed based on its cellular localization. The integration of the transgene into the genome of transgenic plants and the accumulation of the fusion protein were confirmed by using polymerase chain reaction and immunodetection analyses, respectively.
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