[Analysis of the antigenicity of the expression product of fusion antigen epitope genes of FMDV].
OBJECTIVE
From the different recombinants of FMDV antigen epitope, the highest antigenicity one was selected, and that provided research foundation for making researches on FMD plant-based vaccines which could effectively induce cell immune response and be characterized by high eurytopicity.


METHODS
The single-chain DNA of 5 T-cell epitope genes and 2 B-cell epitope genes were synthesized. By gene splicing by overlap extension technique, 5 T-cell epitope genes were fused together into T and 2 B-cell epitope genes were fused together into B. With Ava III and Pst I which produced uniform stickiness terminal, T and B were fused into recombinats in which T and B existed in different position, namely 5'-T-B-T-3', 5'-T-T-B-3' and 5'-B-T-T-3'. With potato X virus expression vector (PVX) every antigen gene (T, B, T-B-T, T-T-B, B-T-T and VP1) was expected to be expressed in leaves of tobacco. RT-PCR analysis and conformed whether all of the antigen genes were transcribed. Indirect ELISA Western Dotblot conformed analysed the antigeni-city of these products.


RESULTS
Every antigen genes were expressed successfully. The different products had different antigenicity, and 5'-T-B-T -3' was highest, 5'-B-T-T-3' was the second, and 5'-T-T-B-3' was lowest, but 5'-T-T-B-3' was higher than VP1, T or B.


CONCLUSIONS
It is possible that gene engineering vaccine against FMDV with high eurytopicity and the induction of cell-immune response could be obtained by the different fusion ways with T and B cell epitopes existing in different position of recombinats, and that could have effect on immune activity of vaccines.
Wang
Dong-mei
DM
Institute of Tropical Biology Sciences, China Academy of Tropical Agriculture Science, Haikou 571101, China. aweeo@tom.com
Shen
Wen-tao
WT
Li
Xiao-ying
XY
Zhou
Peng
P
D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000276:immunology*; D005822:Genetic Vectors / Q000235:genetics; D017863:Potexvirus / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
D000956:Antigens, Viral; D000939:Epitopes; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte