Advanced Biomedical Research

BRIEF REPORT
Year
: 2014  |  Volume : 3  |  Issue : 1  |  Page : 190-

Molecular cloning of Reteplase and its expression in E. coli using tac promoter


Safieh Aghaabdollahian1, Mohammad Rabbani1, Kamran Ghaedi2, Hamid Mir Mohammad Sadeghi1 
1 Department of Pharmaceutical Biotechnology, Isfahan Pharmaceutical Science Research Center, School of Pharmacy and Pharmaceutical Science, Isfahan University of Medical Sciences, Isfahan, Iran
2 Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran

Correspondence Address:
Hamid Mir Mohammad Sadeghi
Department of Pharmaceutical Biotechnology and Isfahan Pharmaceutical Science Research Center, School of Pharmacy and Pharmaceutical Science, Isfahan University of Medical Sciences, Isfahan
Iran

Background and Aims: 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. Materials and 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. Conclusion: 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.


How to cite this article:
Aghaabdollahian S, Rabbani M, Ghaedi K, Sadeghi HM. Molecular cloning of Reteplase and its expression in E. coli using tac promoter .Adv Biomed Res 2014;3:190-190


How to cite this URL:
Aghaabdollahian S, Rabbani M, Ghaedi K, Sadeghi HM. Molecular cloning of Reteplase and its expression in E. coli using tac promoter . Adv Biomed Res [serial online] 2014 [cited 2021 Aug 6 ];3:190-190
Available from: https://www.advbiores.net/article.asp?issn=2277-9175;year=2014;volume=3;issue=1;spage=190;epage=190;aulast=Aghaabdollahian;type=0