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Uchimiya H, Harada H. Transfer of Liposome-Sequestering Plasmid DNA into Daucus carota Protoplasts. Plant Physiol. 1981;68(5):1027-30doi: 10.1104/pp.68.5.1027.
Uchimiya, H., & Harada, H. (1981). Transfer of Liposome-Sequestering Plasmid DNA into Daucus carota Protoplasts. Plant physiology, 68(5), 1027-30. https://doi.org/10.1104/pp.68.5.1027
Uchimiya, H, and Harada, H. "Transfer of Liposome-Sequestering Plasmid DNA into Daucus carota Protoplasts." Plant physiology vol. 68,5 (1981): 1027-30. doi: https://doi.org/10.1104/pp.68.5.1027
Uchimiya H, Harada H. Transfer of Liposome-Sequestering Plasmid DNA into Daucus carota Protoplasts. Plant Physiol. 1981 Nov;68(5):1027-30. doi: 10.1104/pp.68.5.1027. PMID: 16662045; PMCID: PMC426039.
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Plant Physiol. 1981 Nov;68(5):1027-30. doi: 10.1104/pp.68.5.1027.

Transfer of Liposome-Sequestering Plasmid DNA into Daucus carota Protoplasts.

Plant physiology

H Uchimiya, H Harada

Affiliations

  1. Institute of Biological Sciences, University of Tsukuba, Sakura-mura, Ibaraki-ken 305, Japan.

PMID: 16662045 PMCID: PMC426039 DOI: 10.1104/pp.68.5.1027

Abstract

Reverse-phase evaporation lipid vesicles (REV) liposomes, consisting of phosphatidyl choline and stearylamine in 1:3 molar ratio, encapsulated approximately 30% of exogenously supplied recombinant DNA vector, pBR322. The DNA sequestered in REV liposomes was highly tolerant to DNase.A two-step procedure was developed, which involves encapsulation of DNA with liposomes using one-tenth phosphate-buffered saline-0.5 molar mannitol, followed by incubation of liposome-DNA with protoplasts in phosphate buffer-0.5 molar mannitol.About 11% of liposome-encapsulated DNA was transferred into protoplasts, whereas 6% uptake was observed in the control. Although some degradation of incorporated DNA occurred inside protoplasts, 50% of the total radioactivity resolved by 0.8% agarose gel was associated with pBR322 forms in 5-hour incubation. After 20-hour incubation, open circular DNA disappeared completely and maintenance of covalently closed circular DNA was confirmed.

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