Competence for gene transfer by electroporation in a sub-population of protoplasts from uniform carrot cell suspension cultures
✍ Scribed by Robert Bower; Robert G. Birch
- Publisher
- Springer
- Year
- 1990
- Tongue
- English
- Weight
- 404 KB
- Volume
- 9
- Category
- Article
- ISSN
- 0721-7714
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✦ Synopsis
We have investigated the basis for increased transient reporter gene expression following electroporation of protoplasts from uniform carrot cell suspension cultures at increasing DNA concentrations. Use of a combination of histochemical and fluorometric GUS gene assays allowed differentiation between increases due to a higher proportion of expressing protoplasts and increases due to higher expression by each expressing protoplast. A plateau of 20-25% expressing protoplasts was reached by 50 μg ml(-1) DNA but total expression continued to increase in direct proportion to applied DNA concentration up to at least 100 μg ml(-1). This indicates the existence of a subpopulation of protoplasts competent for the uptake and expression of genes by electroporation.