Expression of a single-chain trimer of MHC restricted HBsAg CTL epitope using adenovirus vector containing GFP-report gene
10.3760/cma.j.issn.1003-9279.2009.03.001
- VernacularTitle:采用绿色荧光蛋白腺病毒载体表达HBsAgCTL优势表位肽MHC单链三聚体
- Author:
Xin-Chun CHEN
1
;
Wei-Long LIU
;
Gui-Lin YANG
;
Yong-Jun LIU
;
Xiu-Yun ZHU
;
Hong-Mei ZHANG
;
Bo-Ping ZHOU
;
Lonnie LYBARGER
Author Information
1. 广东省深圳市第三人民医院
- Keywords:
Hepatitis B surface antigens;
Recombinant,genetic;
Adenoviruses,human;
Immunodominant epitopes;
Genes,MHC class Ⅰ;
T-Lymphocytes,cytotoxic;
Fluorescent antibody technique
- From:
Chinese Journal of Experimental and Clinical Virology
2009;23(3):161-164
- CountryChina
- Language:Chinese
-
Abstract:
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.
