Transient expression in leaf mesophyll protoplasts of Arabidopsis thaliana |
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Authors: | Andrea Hoffman Ursula Halfter Peter-Christian Morris |
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Institution: | (1) Institut für Genbiologische Forschung Berlin GmbH, Ihnestraße 63, 1000 Berlin 33, Germany;(2) Present address: Institute des Sciences Végétales, CNRS, Avenue de la Terrasse, 91198 Gif sur Yvette Cedex, France |
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Abstract: | Conditions for maximising transient expression of GUS in leaf mesophyll protoplasts of Arabidopsis thaliana ecotype C24 were investigated. It was found that the factors most influencing expression levels, with optimum levels in parenthesis, were plasmid DNA quantity (100 g per 5 × 105 protoplasts), inclusion of carrier DNA (50 g), PEG pH and amount (pH above 6, and total PEG concentration at least 9% w/w) and the topological form of the DNA. Linearised plasmid DNA with long flanking sequences 3 and 5 to the marker gene yielded the highest levels of GUS expression.Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- GUS
-glucuronidase
- MU
methylumbelliferone
- PEG
polyethylene glycol
- X-gluc
5-bromo-4-chloro-3-indolyl--glucuronic acid |
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Keywords: | Arabidopsis thaliana GUS protoplast transient expression |
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