Expression of a recombinant DNA gene coding for the vesicular stomatitis virus nucleocapsid protein

Abstract

A c[complenetary]DNA clone containing the entire vesicular stomatitis virus nucleocapsid gene was assembled by fusing portions of 2 partial clones. When the cDNA clone was inserted into a new general-purpose eukaryotic expression vector and introduced into appropriate host cells, abundant N-protein synthesis ensued. The expressed protein was indistinguishable from authentic N protein produced during vesicular stomatitis virus infections. The recombinant N protein was recognized by a polyclonal antibody and 2 different monoclonal antibodies and could not be resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from authentic N. Evidently, the recombinant N protein produced in transected [SV40-transformed African green monkey kidney COS] cells rapidly aggregates into high MW complexes in the absence of vesicular stomatitis virus genomic RNA.