Cloning of a Conserved Domain of ORFV- DNA polymerase in the pCDH-CMV-MCS-EF1-cGFP-T2A-Puro Lentiviral Plasmid

Abstract

Received: Revised: Accepted: Published online: June 24, 2018 August 21, 2018 September 06, 2018 March 07, 2019 Contagious ecthyma or Orf is a highly contagious; zoonotic disease and despite the application of some control strategies against it, remains prevalent worldwide. Numerous measures such as anti-viral gene therapies are frequently employed to control viral infections such as ORFV. One way to evaluate these measures is the cloning of the viral target sequence into suitable vectors for the preparation of cell lines expressing subgenomic replicons of the virus. In this study a conserved sequence of ORFVDNA Pol was synthesized and cloned in pCDH-CMV-MCSEF1-cGFP-T2A-Puro lentiviral vector at the upstream of GFP gene. The validity of cloning was confirmed by restriction enzyme digestion and sequencing. Therefore, this recombinant plasmid will be available to produce lentiviral vectors carrying this gene and after infection of Eukaryotic cells with such lentiviral vectors the expression of target gene will be induced. ©2018 PVJ. All rights reserved