Epitope mapping of Aleutian Mink Disease Parvovirus virion protein VP1 and2

Six overlapping fragments of the Aleutian Mink Disease parvo Virus (AMDV) v irion protein VPI and 2 (VP1/ 2) gene were inserted into the expression vec tor pMAL-c2. Four of the clones carried large overlapping fragments coverin g the entire VP1/2 gene. The remaining two clones covered specifically chos en regions within the VP1/2 gene. Using a Western blotting detection system , sera from AMDV-infected mink were tested against the recombinant polypept ides. These studies showed reactions primarily directed against the two AMD V polypeptides ranging from amino acids 297 to 518. Weaker reactions agains t other regions of the VP1/2 were also observed. The small fusion protein d esigned to cover the presumed AMDV VP1/2 loop 4 was purified by affinity ch romatography and used to develop solid-phase immunoassays. Twelve small syn thetic peptides were constructed and used as inhibitors. A peptide covering amino acids S428 to T448 was shown to block the reactivity of a pool of po sitive mink sera, indicating the presence of one dominant linear epitope.