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Plant and Cell Physiology Advance Access originally published online on June 27, 2007
Plant and Cell Physiology 2007 48(8):1098-1107; doi:10.1093/pcp/pcm080
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Transactivation of Protein Expression by Rice HSP101 in Planta and Using Hsp101 as a Selection Marker for Transformation

Ching-Chun Chang*, Pin-Sheng Huang, Huei-Ru Lin and Chun-Hsien Lu

Institute of Biotechnology, National Cheng Kung University, Tainan, Taiwan 701

*Corresponding author: E-mail, chingcc{at}mail.ncku.edu.tw; Fax, +886-6-2766505.


  Abstract

Plant HSP101 has dual activities, first, in conferring thermotolerance, and secondly, in serving as a translational activator. In this study, we introduced Oryza sativa Hsp101 (osHsp101) cDNA into tobacco by Agrobacterium-mediated transformation. Stable integration and expression of the transgene into the tobacco genome was demonstrated by Southern and Western blot analysis. Overexpression of osHSP101 had no noticeable effect on growth or development of the transgenic plants. Homozygous T2 transgenic plants with overexpressed osHSP101 survived heat treatment better than untransformed control plants. In addition, taking advantage of conferring basal thermotolerance by plant HSP101, we were able to demonstrate the feasibility of using osHsp101 as a selection marker and select the transformants under high temperature in tobacco leaf disc transformation mediated by Agrobacterium. Furthermore, transgenic tobacco plants with overexpressed osHSP101 were able to enhance luciferase expression up to 2.9-fold more than untransformed plants in the progeny of reciprocally crossed with omega-luciferase reporter lines.

Keywords: HSP101 - Selection marker - Thermotolerance - Transgenic tobacco - Translational activator

Abbreviations: CaMV, cauliflower mosaic virus; GUS, ß-glucuronidase; HSP101, heat shock protein 101; iLRE, internal light-regulatory element; NAA, naphthalene acetic acid; osHsp101, Oryza sativa Hsp101; TMV, tobacco mosaic virus; UTR, untranslated region

(Received June 4, 2007; Accepted June 14, 2007)
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