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KMID : 0380219940270030190
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Journal of Biochemistry and Molecular Biology
1994 Volume.27 No. 3 p.190 ~ p.195
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Encapsulation of Agrobacterium Ti Plasmid into Liposomes
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Abstract
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Abstract:
@EN Encapsulation of Agrobacterium tumefacient Ti plasmid into liposomes has been studied in order to develop a tool to introduce purified Ti plasmid into plant protoplasts. The efficiency of encapsulation was determined by the radioactivity of
3H-Ti
plasmid and by the fluorescence of octadecyl rhodamine B embedded in the liposome bilayer. The lyophilization-rehydration method encapsulated at least a 20 fold greater amount of the Ti plasmid into liposomes than the reverse-phase evaporation
and
the
Ca2+-EDTA chelation methods. These latter two methods are usually used to encapsulate larger molecules into liposomes. When lyophilization-rehydration method was used, the encapsulation efficiency of the plasmid into liposome ranged from 7 to
30%,
depending upon liposomal lipid compositions. The lipid composition also had great influence on the adhesion of the plasmid on the liposome surface and stability of the encapsulated plasmid. Based on the above mentioned factors,
phosphatidylserine-cholesterol (PS/Chol; to 1 molar ratio) liposome prepared by the lyophilization-rehydration were a good choice for encapsulating Ti plasimid. The optimal ratio of Ti plasmid to lipid was determined to be 15 to 30 ug of Ti
plasmid
per
umol of PS. Under the conditions described, 5 to 8 ug of Ti plasmid was encapsulated into the PS/Chol liposome (umol PS).
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