Transactivation of protein expression by rice HSP101 in planta and using Hsp101 as a selection marker for transformation.
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 T(2) 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.
Chang
Ching-Chun
CC
Institute of Biotechnology, National Cheng Kung University, Tainan, Taiwan 701. chingcc@mail.ncku.edu.tw
Huang
Pin-Sheng
PS
Lin
Huei-Ru
HR
Lu
Chun-Hsien
CH
D015139:Blotting, Southern; D015153:Blotting, Western; D018506:Gene Expression Regulation, Plant; D005822:Genetic Vectors / Q000235:genetics; D006358:Hot Temperature; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D012275:Oryza / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
C115569:HSP101 protein, plant; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D008156:Luciferases