A novel system for producing lentiviral vectors / 生物工程学报
Chinese Journal of Biotechnology
; (12): 958-960, 2007.
Article
en Zh
| WPRIM
| ID: wpr-276180
Biblioteca responsable:
WPRO
ABSTRACT
The aim was to develop a cell culture system capable of producing high titer lentiviral vector stocks with recombinant vaccinia viruses as helpers. BHK21 was co-transfected by three main plasmids containing the transducing plasmid pVECRNA, the packaging plasmid pGAGPOL and the envelop plasmid pVSVG, and thereafter infected with the vaccinia vTF-3 containing bacteriophage T7 RNA polymerase gene using Lipofectamine2000. After 4 days incubation, the culture supernatant of lentiviral vectors was collected and judged by RT-PCR and the Western blot, the results showed that lentiviral vectors were found in the culture supernatant; approximately (11.71 +/- 0.80) x 10(11) copies of lentiviral vector RNA were present per mL of cell culture supernatant, as detected by Real-time PCR; the vector stocks with titers was up to (1.3 +/- 0.18) x 10(8) tu/mL, as detected by flow cytometry , which is one order of magnitude higher than the output of classical manufacture system. These results suggest that the new poxviral/lentiviral hybrid system for efficient lentiviral vector production was initially established. It provides the basis for the future development of industrial application.
Texto completo:
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Índice:
WPRIM
Asunto principal:
Plásmidos
/
Recombinación Genética
/
Virus Vaccinia
/
Proteínas Virales
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ARN Polimerasas Dirigidas por ADN
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Transfección
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Lentivirus
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Técnicas de Cultivo de Célula
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Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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Vectores Genéticos
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2007
Tipo del documento:
Article