title: Assessment the Efficiency of the Constructed Minigenome of Rabies Virus using PV Strain as Helper Virus.
abstract: BACKGROUND
  Rabies is an acute viral disease that causes encephalomyelitis in mammals and human. The only way to prevent this disease is through vaccination before or after exposure. The aim of this study is to evaluate the efficiency of the Pasteur virus (PV) minigenome, using PV strain.
  
  
  METHODS
  Enhanced Green Fluorescent Protein (EGFP) sequence was placed between the designed necessary elements (Hammerhead, HDV ribozyme, 3' Leader, and 5' Trailer sequences), which resemble the rabies virus PV strain (PV2061) genome and anti-genome. These constructs were placed between T7 polymerase promoter and T7 polymerase terminator sequences. The accuracy of the minigenome was confirmed by the expression of EGFP using the helper virus in T7-BHK cell line.
  
  
  RESULTS
  The viral necessary elements of positive and negative sense strands were evaluated for the ability of EGFP expression in the presence of the helper virus. While the positive strand showed background results, no EGFP background was observed in the negative strand application.
  
  
  CONCLUSIONS
  Establishment of minigenome system does not require advanced biosafety levels. Furthermore, using minigenome system eliminates many potential confounding factors that may be present in coding regions of the genome. Use of the minigenome system is easier and more feasible than the full genome rescue of the virus. This study successfully shows the efficiency of the constructed rabies virus minigenome in expression of inserted gene.
authors.affiliation: Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Ajorloo
authors.affiliation: Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. bamdad_t@modares.ac.ir.
authors.forename: Taravat
authors.initials: T
authors.lastname: Bamdad
authors.affiliation: Human Rabies Vaccine Laboratory, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran, WHOCC for Reference and Research on Rabies, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Ali Reza
authors.initials: AR
authors.lastname: Gholami
authors.affiliation: Department of Virology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Keyhan
authors.initials: K
authors.lastname: Azadmanesh
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D016679:Genome, Viral; D049452:Green Fluorescent Proteins / Q000235:genetics; D006378:Helper Viruses / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000235:genetics*; D011818:Rabies / Q000517:prevention & control; D011820:Rabies virus / Q000235:genetics*
chemical_list: D012367:RNA, Viral; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins