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J Clin Microbiol. 1990 May; 28(5): 858-863

Rabies diagnostic reagents prepared from a rabies N gene recombinant expressed in baculovirus.

F L Reid-Sanden, J W Sumner, J S Smith, M Fekadu, J H Shaddock and W J Bellini

Division of Viral Diseases, Center for Infectious Diseases, Atlanta, Georgia 30333.

ABSTRACT

A gene encoding the nucleoprotein (N) of rabies virus was inserted into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus. Recombinant gene expression was controlled by the strong polyhedrin gene promoter. Insect cells (Spodoptera frugiperda) infected by a baculovirus recombinant containing the rabies virus N gene produced abundant amounts of a novel 55-kilodalton protein of a size comparable to that of the rabies virus N protein, as demonstrated by polyacrylamide gel electrophoresis. This new gene product possessed the antigenic and immunogenic properties of native viral N protein, as shown by the ability of the new protein to react in immunoprecipitation and immunofluorescence assays with antirabies antibodies, to serve as a substitute for infectious rabies virus in adsorbing suspensions for diagnostic tests, and to induce high-titered antiserum. The baculovirus expression system provides a safe, convenient, and inexpensive source of rabies virus N protein for the production of both antiserum and adsorbing suspensions for use in rabies diagnoses.

J Clin Microbiol. 1990 May; 28(5): 858-863




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