Expression of Aleutian mink disease parvovirus capsid proteins in a baculovirus expression system for potential diagnostic use.
A 2.3-kb cDNA clone encoding Aleutian mink disease parvovirus (ADV) structural proteins VP1 and VP2 was inserted into the polyhedron gene of Autographa californica nuclear polyhedrosis virus (AcNPV) and expressed by the recombinant virus, AcADV-1, in Spodoptera frugiperda-9 cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western immunoblot analysis (WIA) indicated that synthesis of both VP1 and VP2 was being directed by AcADV-1. Fluorescence microscopic examination of AcADV-1-infected S. frugiperda-9 cells indicated that the recombinant protein was present within the nucleus of the cells, and electron microscopic examination of these cells revealed the presence of small particles 23-25 nm in diameter. Structures resembling empty ADV capsids could be purified on CsCl density gradients, thus indicating that the ADV proteins were self-assembling. The antigenicity of recombinant VP1 and VP2 was evaluated by WIA. Sera collected from 16 mink prior to infection with ADV did not react with VP1 and VP2. Ten sera collected from mink with counter current immunoelectrophoresis (CIE) titers greater than 4 (log2) reacted with VP1 and VP2 in WIA. Two of 6 sera with CIE titers of 4 and 1 of 14 sera with CIE titers < 4 reacted with the recombinant proteins. These results suggest that baculovirus recombinant ADV capsid proteins may be useful as diagnostic antigens.
Department of Microbiology, Immunology and Preventive Medicine, Iowa State University, Ames 50011.
W H
WH
Wu
M E
ME
Bloom
B D
BD
Berry
M J
MJ
McGinley
K B
KB
Platt
D000453:Aleutian Mink Disease / Q000175:diagnosis*; D000454:Aleutian Mink Disease Virus / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D015153:Blotting, Western; D002213:Capsid / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D008854:Microscopy, Electron; D008907:Mink; D009036:Moths; D017924:Nucleopolyhedroviruses / Q000235:genetics; D010957:Plasmids; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification
D011994:Recombinant Proteins