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

Abstract

Border disease virus (BDV) is a serious pathogen of sheep and goats worldwide. The virus causes considerable economic losses in animal husbandry. Therefore, developing effective and genetic-based therapies to control viral infections, such as BDV, has been remarkable over recent years. A common way to evaluate such therapeutic strategies is by cloning the desired fragment into appropriate vectors to develop cell lines expressing it. Due to the required duties of the NS3 gene for BDV proliferation, this gene's HELICc and nucleotide binding site domain was synthesized and cloned in a lentiviral vector upstream of the GFP gene. The cloning accuracy was verified by digestion with restriction enzymes and sequencing. Thus, the BDV-NS3 HELICc and nucleotide binding site carrying plasmid was prepared successfully and will be applied in a second or third lentiviral packaging system for stable expression of the desired gene.