Competence for gene transfer by electroporation in a sub-population of protoplasts from uniform carrot cell suspension cultures

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Abstract
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.