Developing an Antiviral Drug Screening System for Anti-Bovine Viral Diarrhea Virus (BVDV) Therapies

Abstract

Bovine viral diarrhea virus (BVDV) is an economically important animal pathogen affecting cattle. Despite the use of vaccination, test and slaughter practices, BVD remains a serious problem of cattle breeding. This study was conducted in order to develop a cell line that expresses some of BVDV sub-replicons. BVDV-NADL NS3 and 5’UTR were cloned in pWPI-linker B lentiviral plasmid at the upstream of EGFP gene. Consequently, lentiviral vectors containing BVDV-NS3 and BVDV-5’UTR were produced by using the second-generation lentiviral packaging system. By these lentivectors, MDBK cells expressing BVDV-5’UTR and BVDV-NS3 partial fragments were prepared. The efficiency of the infection was evaluated by fluorescence microscopy, western blotting, and RT-PCR. The results indicated that the development of MDBK cell line expressing these transgenes provides a very sensitive antiviral drug screening system for anti-bovine viral diarrhea virus (BVDV) therapies.

Cloning of a BDV-NS3 Conserved Fragment for the Preparation of a Reporter Cell Line

“…The HELICc and nucleotide binding site conserved domain was chosen. Mokhtari et al produced MDBK cells expressed BVDV-5 UTR and BVDV-NS3 using lentiviral vectors (19), used single and multiple siRNA molecules opposed to NS3 CSFV, and showed that viral isolates from different subtypes are inhibited by this method (20). In the present study, for permanent expression of the target genes, the transduction was performed by a lentiviral vector.…”

“…Attenuated live vaccines or recombinant subunits for BDV are not commercially available. Pestivirus contaminants of modified live virus vaccines may cause a serious disease following their inoculation in domestic ruminants (17).…”