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

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...

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Published inApplied microbiology and biotechnology Vol. 44; no. 3/4; pp. 466 - 472
Main Authors Yoshida, K, Kasai, T, Garcia, M.R.C, Sawada, S, Shoji, T, Shimizu, S, Yamazaki, K, Komeda, Y, Shinmyo, A
Format Journal Article
LanguageEnglish
Published Germany 01.12.1995
Subjects
Agrobacterium tumefaciens - genetics
Arabidopsis - genetics
Base Sequence
Biological Sciences
Cells, Cultured
Gene Expression Regulation, Plant
Genes, Reporter
Genetic Vectors
Glucuronidase - biosynthesis
Heat-Shock Proteins - genetics
Hot Temperature
Molecular Sequence Data
Nicotiana - cytology
Nicotiana - metabolism
plant biochemistry
plant breeding
plant genetics
plant physiology
Plant Proteins - biosynthesis
Plant Proteins - genetics
Plants, Toxic
Promoter Regions, Genetic
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Summary: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 HSP 18.2-GUS construct was introduced into BY2 cells by electroporation or Agrobacterium-mediated transformation. Transient expression of the HSP 18.2 promoter in protoplasts was very low regardless of the heat shock. Although expression of the HSP 18.2-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.
ISSN:0175-7598
1432-0614
DOI:10.1007/BF00169945