title: Molecular cloning of Reteplase and its expression in E. coli using tac promoter.
abstract: OBJECTIVE
  This study aimed to clone and express the reteplase cDNA, a thrombolytic agent used for the treatment of acute myocardial infarction and stroke, in E. coli, utilizing tac promoter for its expression.
  
  
  METHODS
  Reteplase cDNA was amplified by polymerase chain reaction (PCR) with designed primers. The product was then cloned into pTZ57R plasmid. The cloned cDNA was digested out and ligated into pGEX-5x-1 expression vector. The presence of the insert was confirmed by restriction digestion. By using 0.2, 0.5 and 1 mM isopropyl beta-D thiogalactopyranoside (IPTG), expression of reteplase was induced in E. coli TOP10 cells and analyzed by SDS-PAGE.
  
  
  RESULTS
  Electrophoresis of PCR product and also double digested recombinant pTZ57R plasmid, also, pGEX-5x-1 vector, showed a 1068bp band of reteplase. SDS-PAGE analysis showed a 60 KDa band of protein product induced with different concentrations of IPTG.
  
  
  CONCLUSIONS
  In the present study, reteplase cDNA was successfully cloned and expressed using tac promoter. This vector will be used for the optimization of the expression of reteplase in E. coli.
authors.affiliation: Department of Pharmaceutical Biotechnology, Isfahan Pharmaceutical Science Research Center, School of Pharmacy and Pharmaceutical Science, Isfahan University of Medical Sciences, Isfahan, I R Iran.
authors.forename: Safieh
authors.initials: S
authors.lastname: Aghaabdollahian
authors.affiliation: Department of Pharmaceutical Biotechnology, Isfahan Pharmaceutical Science Research Center, School of Pharmacy and Pharmaceutical Science, Isfahan University of Medical Sciences, Isfahan, I R Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Rabbani
authors.affiliation: Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, I R Iran.
authors.forename: Kamran
authors.initials: K
authors.lastname: Ghaedi
authors.affiliation: Department of Pharmaceutical Biotechnology, Isfahan Pharmaceutical Science Research Center, School of Pharmacy and Pharmaceutical Science, Isfahan University of Medical Sciences, Isfahan, I R Iran.
authors.forename: Hamid Mir Mohammad
authors.initials: HM
authors.lastname: Sadeghi
keywords: Cloning; reteplase; t-PA; tac promoter