[Expression of a single-chain trimer of MHC restricted HBsAg CTL epitope using adenovirus vector containing GFP-report gene].
OBJECTIVE
To generate a recombinant Adenovirus encoding a GFP (green fluorescent protein)-report gene and a single-chain trimer of MHC restricted HBsAg CTL epitope.


METHODS
An 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.


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 cells demonstrated by western blot assay.


CONCLUSIONS
A recombinant adenovirus expressing HBsAg-SCT and green fluorescent protein report gene has been generated.
Chen
Xin-Chun
XC
Shenzhen Third People's Hospital, Shenzhen 518020, China.
Liu
Wei-Long
WL
Yang
Gui-Lin
GL
Liu
Yong-Jun
YJ
Zhu
Xiu-Yun
XY
Zhang
Hong-Mei
HM
Zhou
Bo-Ping
BP
Lybarger
Lonnie
L
D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D018984:Epitopes, T-Lymphocyte / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006183:H-2 Antigens / Q000235:genetics* / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000378:metabolism; D064532:Histocompatibility Antigen H-2D; D006801:Humans; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D018984:Epitopes, T-Lymphocyte; D006183:H-2 Antigens; D006514:Hepatitis B Surface Antigens; D064532:Histocompatibility Antigen H-2D; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins