Year: 1981
Type: article
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.
Source: PLANT PHYSIOLOGY
DOI: 10.1104/pp.68.5.1027
Authors: Hirofumi Uchimiya, Hiroshi Harada
Institution: University of Tsukuba
Language: English
Subfield: Plant Science
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W2001710064