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Fate of bacterial plasmid DNA during uptake by barley protoplasts
Author(s) -
Hughes Bronwyn G.,
White Fred G.,
Smith Marvin A.
Publication year - 1977
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(77)80355-4
Subject(s) - section (typography) , white (mutation) , graduate students , library science , citation , classics , history , psychology , computer science , biology , genetics , medicine , medical education , gene , operating system
There has been a great deal of excitement generated about the possibility of the genetic transformation of plants in the past few years. Some success has been reported [ 1,2] but the frequency of genetic transformation has been low and the results highly variable [3]. Plant cells devoid of cell walls (protoplasts) would seem to offer an advantage in such studies since they lack the natural barrier to uptake of DNAs, can be grown under selection conditions, and can be regenerated in some cases [4-81. Successful transformation of plants should also depend on efficient introduction of exogenous DNA into plant cells. We have been studying DNA uptake in protoplasts and have found that large amounts of linear bacterial DNA can be taken up. After typical uptake reactions, part of the DNA (20%) was of average genome size while most of the DNA (80%) was depolymerized [9,10]. It seemed that covalently closed circular (CCC) duplex plasmid DNA might be more resistant to depolymerization, and also that it might eventually allow insertion of desirable genes into protoplasts since techniques for gene insertion into plasmids have been developed [ 1 l-141. We have carried out plasmid DNA uptake reactions in barley protoplasts and present the results in this paper.