title: [Establishment of a B16 cell line stably expressing genes encoding HER2 multi-epitope peptides in vivo].
abstract: 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.
authors.affiliation: Department of Immunology, Jilin University, Changchun, China.
authors.forename: He
authors.initials: H
authors.lastname: Li
authors.forename: Ze-Min
authors.initials: ZM
authors.lastname: Xiang
authors.forename: Xiu-Li
authors.initials: XL
authors.lastname: Wu
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Hong-Fei
authors.initials: HF
authors.lastname: Wei
authors.forename: Min
authors.initials: M
authors.lastname: Wan
authors.forename: Li-Ying
authors.initials: LY
authors.lastname: Wang
authors.forename: Yong-Li
authors.initials: YL
authors.lastname: Yu
mesh_terms: 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*
chemical_list: D000939:Epitopes; D049452:Green Fluorescent Proteins; D010446:Peptide Fragments; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins