Enhanced Thermotolerance of E. coli by Expressed OsHsp90 from Rice (Oryza sativa L.) |
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Authors: | Dali Liu Zhenqiang Lu Zijun Mao Shenkui Liu |
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Affiliation: | (1) Key Laboratory of Forest Plant Ecology, Ministry of Education of China, Northeast Forestry University, Harbin, 150040, People’s Republic of China;(2) Key Laboratory of Genetics and Breeding, Academy of Crop Sciences, Heilongjiang University, Harbin, 150080, People’s Republic of China;(3) Key Laboratory of Biochemistry and Molecular Biology, College of Life Sciences, Heilongjiang University, Harbin, 150080, People’s Republic of China;(4) Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, Harbin, 150040, People’s Republic of China; |
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Abstract: | A gene encoding the rice (Oryza sativa L.) 90-kDa heat shock protein (OsHsp90) was introduced into Escherichia coli using the pGEX-6p-3 expression vector with a glutathione-S-transferase (GST) tag to analyze the possible function of this protein under heat stress for the first time. We compared the survivability of E. coli (BL21) cells transformed with a recombinant plasmid containing GST-OsHsp90 fusion protein with control E. coli cells transformed with the plasmid containing GST and the wild type BL21 under heat shock after isopropyl β-d-thiogalactopyranoside induction. Cells expressing GST-OsHsp90 demonstrated thermotolerance at 42, 50, and 70°C, treatments that were more harmful to cells expressing GST and the wild type. Further studies were carried out to analyze the heat-induced characteristics of OsHsp90 at 42, 50, and 70°C in vitro. When cell lysates from E. coli transformants were heated at these heat stresses, expressed GST-OsHsp90 prevented the denaturation of bacterial proteins treated with 42°C heat shocks, and partially prevented that of proteins treated at 50 and 70°C; meanwhile, cells expressing GST-OsHsp90 withstood the duration at 50°C. These results indicate that OsHsp90 functioned as a chaperone, binding to a subset of substrates, and maintained E. coli growth well at high temperatures. |
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