Journal ArticleDOI
Escherichia coli plasmid pBR313 insertion into plant protoplasts and into their nuclei.
Paul F. Lurquin,Clarence I. Kado +1 more
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Cowpea mesophyll protoplasts were shown to bind irreversibly up to 3% input radioactive pBR313 plasmid DNA after 15 min of contact and the presence of donor DNA within the nuclear fraction is most probably not due to an artifactual redistribution of adsorbed plasmID DNA.Abstract:
Cowpea mesophyll protoplasts were shown to bind irreversibly up to 3% input radioactive pBR313 plasmid DNA after 15 min of contact. Maximum uptake occurred in the presence of 5mM ZnSO4 and 5 μg/ml poly-L-ornithine. Under these conditions about one half of the TCA precipitable radioactivity was associated with the nuclear fraction and behaved as linear plasmid molecules. These could not be chased from the protoplasts upon further incubation with unlabeled plasmid DNA. The presence of donor DNA within the nuclear fraction is most probably not due to an artifactual redistribution of adsorbed plasmid DNA. Prolonged incubation periods resulted in extensive degradation of plasmid in the incubation medium but little degradation occurred in the protoplasts. The donor DNA was not covalently associated with the protoplast nuclear DNA.read more
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Rapid procedure for detection and isolation of large and small plasmids.
C I Kado,Shih-Tung Liu +1 more
TL;DR: The method utilized the molecular characteristics of covalently closed circular deoxyribonucleic acid (DNA) released from cells under conditions that denature chromosomal DNA by using alkaline sodium dodecyl sulfate (pH 12.6) at elevated temperatures.
Journal ArticleDOI
Direct gene transfer to plants.
Jerzy Paszkowski,Raymond D. Shillito,Michael W. Saul,Vaclav Mandak,Thomas Hohn,Barbara Hohn,Ingo Potrykus +6 more
TL;DR: Plants regenerated from transformed cell lines were phenotypically normal and fertile, and they maintained and expressed the foreign gene throughout the development of vegetative and generative organs.
Journal ArticleDOI
Transformation of the green alga Chlamydomonas reinhardii with yeast DNA
TL;DR: The first successful nuclear transformation of C. reinhardii is described, where a plasmid carrying the yeast arg4 locus and a yeast replication origin has been used to transform a C. Reinhardii cell wall-deficient, arginine-requiring mutant.
Journal ArticleDOI
Gene transfer in cereals
TL;DR: By means of direct interaction of cereal protoplast with plasmids, coupled with improved procedures for the regeneration of plants from their protoplasts, gene transfer in the cereals is becoming established at the frontiers of recombinant DNA technology.
Journal ArticleDOI
Direct gene transferState of the Art and Future Potential
TL;DR: Host range limitations were not expected and also not found in transformation experiments with protoplasts from a series of herbaceous dicots and a graminaceous monocot, and co-transformation experiments with a selectable and a non-selectable gene on separate plasmids yielded high levels of cotransfer of the non- selecting gene.
References
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Journal ArticleDOI
Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis
Journal ArticleDOI
Construction of a colicin E1-R factor composite plasmid in vitro: means for amplification of deoxyribonucleic acid.
T Tanaka,Bernard Weisblum +1 more
TL;DR: The continued replication in the presence of chloramphenicol suggests that the replicator of the colicin E1 factor is functional in the composite plasmid, and conferred on the host cell resistance to streptomycin, sulfonamide-cesium chloride solution.
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Molecular structure of bacterial plasmids.
TL;DR: The present work focuses on the development of a model for rFactor Evolution by analyzing the role of F and F' Sex Factors in the evolution of R-Factor Evolution.
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Effect of growth conditions on the formation of the relaxation complex of supercoiled ColE1 deoxyribonucleic acid and protein in Escherichia coli.
TL;DR: Inhibition of protein synthesis in log cultures by the addition of chloramphenicol or amino acid starvation allows ColE1 DNA to continue replicating long after chromosomal replication has ceased.
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Transposable genetic elements and plasmid evolution
TL;DR: Transposable elements of DNA that are structurally defined and genetically discrete units seem to have an important role in the evolution of bacterial plasmids and serve as novel biological switches capable of turning on and off the expression of nearby genes as a consequence of their insertion into or excision from plasmid genomes.