Construction and characterization of an attenuated bovine herpesvirus type 1 (BHV-1) recombinant virus

A recombinant bovine herpesvirus type 1 (BHV-1) virus (Gal-TK) has been constructed. The Gal-TK virus contains a chimeric reporter/marker gene coding for bacterial beta-galactosidase (beta-gal gene) that was inserted stably within the viral TK gene. This resulted in inactivation of the TK gene. The beta-gal gene is under the regulation of a strong, human cytomegalovirus-immediate early (HCMV-IE) promoter and is expressed as an authentic viral-coded gene. Even though the one-step growth kinetics of the recombinant and parent viruses were similar, the recombinant virus yielded less than the parent virus on Madin-Darby bovine kidney cells. After intranasal inoculation, the engineered virus was virtually avirulent for colostrum-deprived new-born calves. Similar to the parent virus, the recombinant virus replicated in the upper respiratory tract of calves, but the amount of progeny viruses produced was reduced significantly. The progeny viruses recovered from nasal swabs of animals inoculated with the recombinant and the Cooper strains of BHV-1 were easily distinguishable based on the beta-gal marker.