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Published on: June 19, 2013
[Preparation and transfection of GFP plasmid DNA/cationic liposome complex]
Li Liu1, Wen-xiang Jia, Yong-qiu Mao
1Department of Microbiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Objective:
This work was directed at obtaining a better gene carrier to improve the effects of gene delivery.
Methods:
Cationic liposomes made from cholesterol, lecithin and Eighteenth Amic by reverse evaporation technique were used for encapsulating plasmid DNA containing gfp gene. The DNA/liposome complexes differed in quantity of plasmid DNA. The sizes of complexes and the efficiency of encapsulation were detected. MTT assay was used to measure the cytotoxicities of complexes. The efficiency of transfection into COS1 cells was shown by observation of green fluorescent and measurement of fluorescent intensity.
Results:
The average size of complexes was 562 nm, the encapsulating efficiency of DNA in microspheres reached 55%-65%. These DNA/Cationic liposome complexes could be transfected into mammalian cells, but they were different from each other in efficiency of transfection. The cytotoxicities of these complexes varied with the concentration of DNA in complexes, the quantities of complexes and the time of treatment by complexes.
Conclusion:
DNA/Cationic liposome complexes prepared by reverse evaporation technique could be applied in DNA delivery system.
