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. 1981 Nov;68(5):1027–1030. doi: 10.1104/pp.68.5.1027

Transfer of Liposome-Sequestering Plasmid DNA into Daucus carota Protoplasts

Hirofumi Uchimiya 1, Hiroshi Harada 1
PMCID: PMC426039  PMID: 16662045

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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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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