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Updated: Feb 13, 2026
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Transient Gene Expression in Tobacco using Gibson Assembly and the Gene Gun
Published on: April 18, 2014
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Expression of T7-based constructs in tobacco cells.
Hyukho Sheen1, K Andrew White1
1Department of Biology, York University, 4700 Keele Street, Toronto, Ontario, M3J 1P3, Canada.
Biochemical and Biophysical Research Communications
|March 21, 2018
Summary
Bacteriophage T7 RNA polymerase (T7-Pol) shows limited success for protein expression in plant cells. Nuclear T7 transcription failed, but cytosolic transcription with a viral enhancer enabled protein expression in tobacco.
Area of Science:
- Plant molecular biology
- Biotechnology
- Gene expression systems
Background:
- Bacteriophage T7 promoter and RNA polymerase (T7-Pol) are common for recombinant protein production.
- Conflicting results exist regarding T7-Pol efficacy for gene expression in plants.
Purpose of the Study:
- To resolve discrepancies in plant T7-Pol expression systems.
- To evaluate green fluorescent protein (GFP) expression from T7 constructs in tobacco.
Main Methods:
- Assessed GFP expression in tobacco protoplasts using T7 constructs.
- Utilized agroinfiltration for transient expression studies in whole plants.
- Investigated effects of nuclear vs. cytosolic T7-Pol and viral translation enhancers.
Main Results:
- Nuclear T7 transcription did not yield GFP in plant protoplasts.
- Cytosolic T7 transcription with a viral enhancer allowed GFP translation.
- Nuclear T7 transcripts and modified export strategies failed to improve expression in plants.
Conclusions:
- Nuclear T7 construct expression is not viable in tobacco cells.
- Cytosolic transcription offers a feasible alternative for direct RNA overexpression in the plant cytosol.
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