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Clinical and Diagnostic Laboratory Immunology, August 2005, p. 904-909, Vol. 12, No. 8
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.8.904-909.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Production of a Recombinant Major Inner Capsid Protein for Serological Detection of Epizootic Hemorrhagic Disease Virus

Lizhong Luo* and Marta I. Sabara

Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, Canadian Science Centre for Human and Animal Health, Winnipeg, Manitoba, Canada

Received 11 March 2005/ Returned for modification 19 April 2005/ Accepted 4 May 2005

Constructs of the major core protein, designated VP7, from epizootic hemorrhagic disease virus (EHDV) type 1 were made by amino- or carboxyl-terminal fusion of a six-histidine residue tag to the VP7-1 gene. The resulting fusion proteins were produced in a baculovirus expression system and purified by a rapid, one-step procedure using nickel-nitrilotriacetic acid technology. A high level of VP7-1 protein expression was detected with the N-terminal six-histidine tag fusion construct and was comparable to the level of expression observed with an untagged VP7-1 Bam construct. In contrast, the inclusion of a six-histidine tag at the C terminus adversely affected protein expression. The antigenicity of the N-terminal six-histidine tag EHDV VP7-1 product was identical to that observed with the native virus antigen and untagged EHDV VP7-1 recombinant protein, as determined by reactivity with EHDV specific antibodies in an enzyme-linked immunosorbent assay (ELISA) and Western blot. The high production and purity levels that can be attained for the N-terminal six-histidine tag VP7-1 protein and its reactivity with EHDV-specific sera in a competitive ELISA make it a suitable assay reagent.

* Corresponding author. Mailing address: Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, 1015 Arlington St., Winnipeg, Manitoba R3E 3M4, Canada. Phone: (204) 789-2149. Fax: (204) 789-2038. E-mail: luol{at}inspection.gc.ca.

Clinical and Diagnostic Laboratory Immunology, August 2005, p. 904-909, Vol. 12, No. 8
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.8.904-909.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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