Cloning of BHV-UL25 Conserved Fragment in a Lentiviral Transfer Plasmid for the Preparation of a Monitoring Cell Line

Abstract

: Bovine herpesvirus-1 (BHV-1) is the causative agent of a domestic and wild cow disease namely infectious bovine rhinotracheitis (IBR). Severe economic damage due to IBR is possible to occur and vaccines are not completely effective against the disease. Therefore, over the recent years, the development of effective and genetic-based therapies to control viral infections, such as IBR has been remarkable. A common way to evaluate such therapeutic strategies is cloning of the viral target sequence into appropriate vectors for the preparation of cell lines expressing viral subgenomic replicons. Due to the required duties of UL25 gene, serine protease substrate domain of this gene was cloned in pCDH-CMV-MCS-EF1-cGFP-T2A-Puro lentiviral vector at the upstream of GFP gene. The cloning accuracy was verified by restriction of enzyme digestion and sequencing. Thus, this recombinant plasmid will be available to produce lentiviral vectors with the desired gene; after infection of eukaryotic cells with such lentiviral vectors the target gene will be expressed.