A Recombinant Classical Swine Fever Virus Stably Expresses a Marker Gene

The gene coding for bacterial chloramphenicol acetyltransferase (CAT) was inserted in frame into the viral N(pro) gene of the full-length cDNA clone pA187-1 of the classical swine fever virus (CSFV) strain Alfort/187. RNA transcribed in vitro from the resulting plasmid was transfected into SK-6 porcine kidney cells. Infectious progeny virus vA187-CAT recovered from transfected cells had growth characteristics indistinguishable from those of parental virus vA187-1. In cells infected with vA187-CAT the predicted fusion protein, CAT-N(pro), was detected, and it retained the enzymatic activities of both CAT and N(pro). The CAT gene remained stably inserted in the viral genome after 10 virus passages. Thus, marker virus vA187-CAT represents a useful tool for quantitative analysis of viral replication and gene expression.