Recombinant swinepox virus expressing beta-galactosidase: investigation of viral host range and gene expression levels in cell culture.
Swinepox virus (SPV) has been proposed as a potential vector for generating recombinant vaccines for swine. However, little is known about important aspects of SPV biology, such as the functionality of SPV promoters or the host range of SPV. Using a transient expression assay, well-characterized vaccinia virus promoters were shown to be active in cells infected with SPV. A recombinant SPV expressing beta-galactosidase (beta-gal) was constructed and characterized. The E. coli LacZ gene was placed under the control of a strong vaccinia synthetic early/late promoter and was inserted by homologous recombination in a noncoding region of the SPV genome. The recombinant SPV expressing beta-gal was used to characterize the host range of the virus by measuring protein expression and virus production in different cell lines. In general, SPV expressed more protein and grew more efficiently than vaccinia virus in porcine cell lines. Surprisingly, the recombinant SPV was able to infect and replicate in several cell lines of nonswine origin. The virus directed regulated early and late gene expression of beta-gal in those cells and formed blue plaques in cell monolayers in the presence of X-gal. Upon infection with the recombinant SPV, there was a significant level of viral replication, and the virus can be serially passaged in some nonswine cell lines. The data presented suggest that despite the strict host tropism of SPV, the virus exhibits a relatively broad host range in cell culture.
Bárcena
J
J
Centro de Investigación en Sanidad Animal-I.N.I.A., Valdeolmos, Madrid, Spain.
Blasco
R
R
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D016254:Mutagenesis, Insertional; D011817:Rabbits; D011995:Recombination, Genetic; D018148:Suipoxvirus / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D013552:Swine; D014616:Vaccinia virus / Q000502:physiology; D014779:Virus Replication; D001616:beta-Galactosidase / Q000235:genetics* / Q000302:isolation & purification
D001616:beta-Galactosidase