Competence for gene transfer by electroporation in a sub-population of protoplasts from uniform carrot cell suspension cultures.
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.
Department of Botany, The University of Queensland, 4072, Australia.
R
R
Bower
R G
RG
Birch