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


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Li
He
H
Department of Immunology, Jilin University, Changchun, China.
Xiang
Ze-Min
ZM
Wu
Xiu-Li
XL
Wang
Li
L
Wei
Hong-Fei
HF
Wan
Min
M
Wang
Li-Ying
LY
Yu
Yong-Li
YL
D000818:Animals; D045744:Cell Line, Tumor; D000939:Epitopes / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008546:Melanoma, Experimental / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
D000939:Epitopes; D049452:Green Fluorescent Proteins; D010446:Peptide Fragments; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins