title: A hepadnavirus regulatory element enhances expression of a type 2 bovine viral diarrhea virus E2 protein from a bovine herpesvirus 1 vector.
abstract: Recently, the possibility of using virus vectors to immunize cattle against selected bovine viral diarrhea virus (BVDV) genes has gained widespread interest. However, when we attempted to express the E2 protein from type 2 (890 strain) BVDV in a bovine herpesvirus 1 (BHV1) vector, we observed that expression was poor. This often happens when genes from a cytoplasmic virus are expressed in the cell nucleus. To counter this effect, we attempted to enhance expression by a strategy employed by viruses. RNAs of retroviruses and hepadnaviruses contain cis-acting elements that facilitate expression of RNAs that otherwise are degraded or retained within the nucleus. In Mason-Pfizer monkey virus, the required RNA sequence element is known as a constitutive transport element (CTE). A related element from woodchuck hepatitis virus is known as the woodchuck posttranscriptional regulatory element (WPRE). We tested the ability of the CTE, the WPRE, and introns to enhance expression of E2. All three elements stimulated expression of E2 from plasmids. The combination of the WPRE and an intron yielded the highest level of E2 expression in plasmids. However, when E2 was expressed from a BHV1 vector, the presence of an intron was inhibitory. In contrast, the WPRE was very efficient at stimulating E2 expression from a BHV1 vector. This result represents the first expression of a type 2 BVDV E2 protein from a mammalian virus vector and raises the possibility that the WPRE may provide a general method of enhancing foreign gene expression from BHV1 and other herpesvirus vectors.
authors.affiliation: Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Lingshu
authors.initials: L
authors.lastname: Wang
authors.forename: Sreekumar
authors.initials: S
authors.lastname: Menon
authors.forename: Steven R
authors.initials: SR
authors.lastname: Bolin
authors.forename: Leonard J
authors.initials: LJ
authors.lastname: Bello
mesh_terms: D000818:Animals; D002417:Cattle; D030242:Diarrhea Virus 2, Bovine Viral / Q000235:genetics*; D005822:Genetic Vectors; D016679:Genome, Viral; D015613:Hepadnaviridae / Q000235:genetics*; D007242:Herpesvirus 1, Bovine / Q000235:genetics*; D016366:Open Reading Frames; D010957:Plasmids; D011995:Recombination, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D014764:Viral Proteins