1.Expression of a single-chain trimer of MHC restricted HBsAg CTL epitope using adenovirus vector containing GFP-report gene
Xin-Chun CHEN ; Wei-Long LIU ; Gui-Lin YANG ; Yong-Jun LIU ; Xiu-Yun ZHU ; Hong-Mei ZHANG ; Bo-Ping ZHOU ; Lonnie LYBARGER
Chinese Journal of Experimental and Clinical Virology 2009;23(3):161-164
Objective To generate a recombinant Adenovh'us encoding a GFP (green fluorescent protein)-report gene and a single-chain trimer of MHC restricted HBsAg CTL epitope. Methods An oligonucleotide encoding H-2Ld restricted HBsAg CTL epitopo was synthesized and fused with H-2Ld 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 transfeeted into Ad293 cells for packaging and amplification of recombinant adenovirus encoding HBsAg-SCT. Results HBsAg-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 ceils demonstrated by western blot assay. Conclusion A recombinant adenovirus expressing HBsAg-SCT and green fluorescent protein report gene has been generated.
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