| Autor: |
He, Li, Ze-Min, Xiang, Xiu-Li, Wu, Li, Wang, Hong-Fei, Wei, Min, Wan, Li-Ying, Wang, Yong-Li, Yu |
| Rok vydání: |
2009 |
| Předmět: |
|
| Zdroj: |
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology. 25(1) |
| ISSN: |
1007-8738 |
| Popis: |
To establish a B16 cell line stably expressing genes encoding HER2 multi-epitope peptides in vivo.Eukaryotic expressing vector pcDNA3-GFP-HER2 was constructed by molecular cloning technique and transfected into B16 cells mediated by cationic liposome. After screened with G418, the transfected cells were passaged in vivo. The GFP-HER2 positive cells were isolated from tumor burdening mice by flow cytometry (FCM) sorting, and then monoclonized in the absence of G418 to obtain a B16 cell line stably expressing genes encoding HER2 multi-epitope peptides in vivo. This cell line was then identified after being passaged in vivo.Restriction endonulease analysis and DNA sequencing showed that the pcDNA3-GFP-HER2 was constructed and a B16 cell line stably expressing genes encoding HER2 multi-epitope peptides in vivo was obtained successfully. The GFP-HER2 positive proportion maintained higher than 90% after being passaged in vivo.A B16 cell line stably expressing genes encoding HER2 multi-epitope peptides is established successfully, which would provide a method to establish other cell lines stably expressing exogenous genes. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
|
