STRUCTURAL AND ANTIGENIC ANALYSIS OF THE NUCLEOPROTEIN OF BOVINE EPHEMERAL FEVER RHABDOVIRUS

The nucleotide sequence of the bovine ephemeral fever virus (BEFV) gen ome has been determined from the 3' terminus to the end of the nucleop rotein (N) gene. The 3' leader sequence comprises 50 nucleotides and s hares a common terminal three nucleotides (3'-UGC-) and a downstream U -rich domain with vesicular stomatitis virus (VSV) and rabies virus. T he N gene comprises 1328 nucleotides from the transcription initiation consensus sequence (AACAGG) to the conserved transcription terminatio n-poly(A) sequence [CATG(A)(7)] and encodes a polypeptide of 431 amino acids with an estimated M(r) of 49159 and a pI of 5.4. The deduced am ino acid sequence of the BEFV N protein is similar to those of other m ammalian rhabdoviruses and is more closely related in sequence to vesi culoviruses (VSV Indiana and New Jersey, Piry, Chandipura) than to lys saviruses (rabies and Mokola). An almost full-length clone, 1301 bp in length, of the BEFV N gene and clones derived from 5'-terminal (559 b p) and 3'-terminal (742 bp) fragments were expressed in Escherichia co li as glutathione-S-transferase fusion proteins. A panel of 12 BEFV N protein-specific monoclonal antibodies was shown to react in immunoblo ts with fusion proteins containing the almost full-length N protein an d the C-terminal fragment, but not the N-terminal fragment. Two of the se antibodies also reacted with baculovirus-expressed rabies virus N p rotein. Polyclonal mouse ascitic fluids derived from BEFV, rabies viru s and several other related viruses were also shown to crossreact in i mmunoblots with purified preparations of rabies virus and BEFV N prote ins.