ENCEPHALITIS-VIRUS E-PROTEIN EXPRESSED IN RECOMBINANT ESCHERICHIA-COLI SYSTEM

In order to localize denaturation-resistant neutralizing eptiope(s) in the C-terminal 180 amino acids of Japanese encephalitis (JE) virus E- protein, four recombinant clones encoding different or overlapping nuc leotide sequences were constructed by PCR from a recombinant plasmid p S22. The amplified fragments were cloned into PCR 1000 vector, and the n transferred into Escherichia coli expression vector pRIT2T. The inse rted genes were expressed as fusion proteins with protein-A and examin ed for their antigenicity and immunogenicity by Western blotting and m ouse immunization, respectively. Among the four recombinant fusion pro teins, the highest neutralizing antibody titre was obtained by the one expressed by the recombinant clone pRIT2T-B3, which carried the codin g sequence of amino acid number 373-399 of JE virus E protein. The res ults indicated that this short region of 27 amino acids sequence near the C-terminal of JE virus E protein possesses neutralizing epitope(s) . These data should assist in the design of an efficient subunit vacci ne against JE virus infection in future.