[Establishment of a B16 cell line stably expressing genes encoding HER2 multi-epitope peptides in vivo]

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.