OBJECTIVETo generate a recombinant Adenovirus encoding a GFP (green fluorescent protein)-report gene and a single-chain trimer of MHC restricted HBsAg CTL epitope.

METHODSAn oligonucleotide encoding H-2L(d) restricted HBsAg CTL epitope was synthesized and fused with H-2L(d) DNA molecule to construct the eukaryotic expression vector carrying the HBsAg-SCT gene. The HBsAg-SCT gene was subcloned into a GFP adenovirus expression vector,which was transfected into Ad293 cells for packaging and amplification of recombinant adenovirus encoding HBsAg-SCT.

RESULTSHBsAg-SCT has been cloned into an adenovirus vector encoding GFP report gene successfully as confirmed by double enzyme digestion and direct sequencing. HBsAg-SCT was expressed by infected Ad293 cells demonstrated by western blot assay.

CONCLUSIONA recombinant adenovirus expressing HBsAg-SCT and green fluorescent protein report gene has been generated.

Adenoviridae ; genetics ; metabolism ; Animals ; Cell Line ; Epitopes, T-Lymphocyte ; genetics ; metabolism ; Gene Expression ; Genes, Reporter ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; H-2 Antigens ; genetics ; metabolism ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Histocompatibility Antigen H-2D ; Humans ; Mice ; Recombinant Fusion Proteins ; genetics ; metabolism