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- W2347251913 abstract "Objective: To explore the impact of different conformations of DNA vaccine on expression levels in vitro and in vivo.Methods: The pcDNA3.1-EGFP and pDRVISV1.0-Fluc plasmids were constructed respectively.The effects of three existing topological forms(including supercoiled,linear,and closed-loop nicked forms) and the repeatedly freezing and thawing condition on the expression efficiency were analyzed.For the former,liposome was used to mediate the transfection of 293FT cells and the expression of EGFP was detected using flow cytometer;for the latter,the in vivo imaging technique was used to monitor the expression of luciferase after inoculating the BALB / c mice.Results: For pcDNA3.1-EGFP plasmids,after the transfection of 293FT cells for 48 hours,the flow cytometry showed the expression rates and fluorescence intensity of EGFP expressed in the plasmids of the four treatment groups varied as follows: freshly prepared plasmids(89.76%,2682.68) repeatedly freeze-thaw processing plasmids(76.35%,2313.48) closed-loop nicked plasmids(68.45%,1245.95) linear plasmids(38.28%,929.5).After pDRVISV1.0-Fluc plasmids were immunized into mice,the luciferase fluorescence signal intensity was observed for 28 days using in vivo imaging technology.The results showed that the fluorescence were observed at 0.2 day after immunization,after which the intensity began to continuously increase to a plateau at Day 5 and maintained to Day 18.Significant differences were observed in fluorescence intensity between each treatment group,in an order of freshly prepared plasmids(108.6) repeated freezing and thawing processing plasmids(108.4) linear plasmids(108.3) nicked plasmids(108.0).The in vivo and in vitro expression was the highest for the freshly prepared plasmids which were mainly composed of supercoiled DNA.Conclusion: Higher proportion of supercoiled plasmids may lead to the higher transfection efficiency and expression level,and increasing the proportion of supercoiled plasmids is the key to quality control for DNA vaccine." @default.
- W2347251913 created "2016-06-24" @default.
- W2347251913 creator A5027810140 @default.
- W2347251913 date "2013-01-01" @default.
- W2347251913 modified "2023-09-27" @default.
- W2347251913 title "Study on differences in expression levels in vitro and in vivo between different conformational forms of DNA vaccine" @default.
- W2347251913 hasPublicationYear "2013" @default.
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