Heat-inducible expression system for a foreign gene in cultured tobacco cells using the HSP18.2 promoter of Arabidopsis thaliana

Applied Microbiology and Biotechnology
K YoshidaA Shinmyo

Abstract

A system for the controlled expression of a foreign gene in cultured tobacco cells (Nicotiana tabacum, BY2) by temperature shift was constructed. A 925-base-pair (bp) DNA fragment containing the 5'-flanking region of a low-molecular-mass heat-shock protein gene (HSP18.2) of Arabidopsis thaliana was inserted upstream of the beta-glucuronidase reporter gene (GUS). The resulting HSP18.2-GUS construct was introduced into BY2 cells by electroporation or Agrobacterium-mediated transformation. Transient expression of the HSP18.2 promoter in protoplasts was very low regardless of the heat shock. Although expression of the HSP18.2-GUS chimeric gene in the stable transformants of BY2 was hardly detected in culture at 25 degrees C, the expression increased rapidly on the transcriptional level when the incubation temperature was shifted to 35-37 degrees C. The optimal temperature for heat-shock induction was 37 degrees C. After a 2-h incubation at 37 degrees C, GUS activity was about 1000-fold greater than that before heat shock. The amount of GUS mRNA was maximum 2 h after heat shock, and then decreased gradually.

Citations

Apr 7, 1999·Biotechnology and Bioengineering·A ShinmyoK Yoshida
Sep 8, 2006·Applied Microbiology and Biotechnology·Kung-Ta LeeTakashi Yamakawa
Oct 20, 2005·Journal of Bioscience and Bioengineering·S NagayaA Shinmyo
Oct 20, 2005·Journal of Bioscience and Bioengineering·K Yoshida, A Shinmyo
May 23, 2009·Biotechnology Advances·Giandomenico Corrado, Marianthi Karali
Mar 5, 2004·The Plant Journal : for Cell and Molecular Biology·Jérôme JoubèsLieven De Veylder
Jan 31, 2006·The Plant Journal : for Cell and Molecular Biology·Ram DixitSimon Gilroy
Jan 30, 2008·Current Protocols in Cell Biology·Federica BrandizziChris Hawes
May 24, 2005·American Journal of Physiology. Heart and Circulatory Physiology·Sylvain RoyNikolaos Stergiopulos
Jul 9, 2014·Biological Chemistry·Frederik FadenNico Dissmeyer
Oct 20, 2005·Journal of Bioscience and Bioengineering·Tomoko DansakoAtsuhiko Shinmyo

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