COMPLETE NUCLEOTIDE-SEQUENCE OF THE HERPESVIRUS-SIMIAE GLYCOPROTEIN-GGENE AND ITS EXPRESSION AS AN IMMUNOGENIC FUSION PROTEIN IN BACTERIA

The nucleotide sequence of a 2384 bp portion within the unique short ( U-s) region of the herpesvirus simiae (simian herpes B virus; SHBV) ge nome is presented. A partial and a complete open reading frame (ORF) w ere found within this nucleotide sequence. The partial ORF encodes the C terminus (147 amino acids) of a protein kinase which is highly cons erved in the herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) an d simian agent 8 (SA8) U-s regions. The complete ORF is located 3' to the partial ORF within the 2384 bp sequence and encodes a 593 amino ac id glycoprotein which appears to be closely related to the SA8 glycopr otein G (gG), but shares little amino acid similarity with gG of HSV-1 and -2. However, the complete ORF shares certain features conserved a mong most alphaherpesvirus gGs, notably three highly conserved cystein e residues and an adjacent N-glycosylation site. Therefore, it was con cluded that this complete ORF encodes the SHBV gG. The 358 amino acid C-terminal portion of SHBV gG was expressed in Escherichia coli as a f usion protein and this was detected by immunoblotting with sera from c ynomolgus monkeys which were either experimentally or naturally infect ed with SHBV. The purified fusion protein was inoculated into rabbits to raise an antiserum which recognized a number of apparently SHBV gG- specific protein bands in extracts from SHBV-infected simian cells.