COMPARATIVE SEQUENCE-ANALYSIS OF A POLYMORPHIC REGION OF THE SPIKE GLYCOPROTEIN S1 SUBUNIT OF ENTERIC BOVINE CORONAVIRUS ISOLATES

Complementary oligonucleotide primers which flank a 1146-nucleotide gene fragment (S1B: nt 1185 to 2333) encompassing a polymorphic region (nt 1368 to 1776) of the S1 subunit of bovine coronavirus spike glycoprotein were used for enzymatic amplification by PCR. We chose four clinical isolates, recovered from cases of epidemic diarrhea in neonatal calves in Quebec dairy herds between 1987-1990, to specifically amplify and analyze their sequences in the selected genomic area. Nucleotide sequence analysis of the four clinical isolates indicated that their S1B gene fragments were highly conserved. We also compared the S1B gene sequences of the Quebec BCV isolates to the published corresponding sequences from BCV-L9 [37], BCV-MEB [1], and BCV-F15 [3] reference strains. A high degree of similarity was demonstrated for all viruses, no deletions or insertions were observed, and the only variations that were identified consisted of nucleotide substitutions. The differing nucleotides and amino acids (aa) were not distributed randomly over the entire sequence but rather were clustered in the polymorphic region. Of these, four sporadic aa changes were located in antigenic domain II (aa residues 517 to 720) of S1. This correlates with varied antigenicity observed among the BCV Quebec isolates when reacting with MAbs directed against the S glycoprotein of the Mebus strain. The other mutations seem to be fixed in all Quebec isolates.