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Heat-inducible expression system for a foreign gene in cultured tobacco cells using the HSP18.2 promoter of Arabidopsis thaliana.
- Source :
- Applied Microbiology & Biotechnology; 1995, Vol. 44 Issue 3/4, p466-472, 7p
- Publication Year :
- 1995
-
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 β-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°C, the expression increased rapidly on the transcriptional level when the incubation temperature was shifted to 35-37°C. The optimal temperature for heat-shock induction was 37°C. After a 2-h incubation at 37°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. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 01757598
- Volume :
- 44
- Issue :
- 3/4
- Database :
- Complementary Index
- Journal :
- Applied Microbiology & Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 72380056
- Full Text :
- https://doi.org/10.1007/BF00169945