title: Liposome-mediated delivery of DNA to carrot protoplasts.
abstract: The encapsulation of DNA within liposomes and subsequent fusion of the liposomes with carrot (Daucus carota L.) protoplasts were examined to determine optimum conditions for effective liposome-mediated delivery of DNA to protoplasts. Escherichia coli [(3)H]DNA could be encapsulated with 50% efficiency using encapsulation volumes as low as 0.5 ml. Incorporation of liposome-encapsulated [(3)H]DNA by carrot protoplasts increased linearly for 2.5 h, and increasing the ratio of protoplasts to liposomes increased the total amount of radioactive label incorporated within the protoplasts. Liposome-mediated incorporation of [(3)H]DNA by protoplasts increased over a range of polyethylene glycol concentrations up to 20%, but Ca(2+) did not increase liposome-mediated incorporation when present in the liposome-protoplast incubation mixture. Optimum incorporation was observed when the pH of the liposome-protoplast incubation medium was decreased to 4.8. Encapsulation experiments using DNA of the plasmid pBR322 indicated that an average of 200-1,000 intact copies of pBR322 were sequestered within each nucleus after liposome delivery.
authors.affiliation: Cell Culture and Nitrogen Fixation Laboratory, Plant Physiology Institute, Agricultural Research, Science and Education Administration, U.S. Department of Agriculture, 20705, Beltsville, MD, USA.
authors.forename: B F
authors.initials: BF
authors.lastname: Matthews
authors.forename: D E
authors.initials: DE
authors.lastname: Cress