ArticleAdvanced biomedical research2014
Molecular cloning of Reteplase and its expression in E. coli using tac promoter.
Article in Advanced biomedical research, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 19 citations in OpenAlex.
- Response Surface Methodology to Optimize the Expression Efficiency of Recombinant Reteplase.Iranian journal of biotechnology · 2023Article
- Construction of Strong Promoters by Assembling Sigma Factor Binding Motifs.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Improving the solubility, activity, and stability of reteplase usingResearch in pharmaceutical sciences · 2019Article
- Reteplase: Structure, Function, and Production.Advanced biomedical research · 2019Review
- Evaluation of efficacy and safety of Reteplase and Alteplase in the treatment of hyper-acute cerebral infarction.Bioscience reports · 2018Article
- Article
- Chromosome engineering of Escherichia coli for constitutive production of salvianic acid A.Microbial cell factories · 2017Article
- High-level expression of a novel recombinant human plasminogen activator (rhPA) in the milk of transgenic rabbits and its thrombolytic bioactivity in vitro.Molecular biology reports · 2016Article
- Comparison of Three Escherichia coli Strains in Recombinant Production of Reteplase.Avicenna journal of medical biotechnologyArticle
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
BACKGROUND AND
aimsThis 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
methodsReteplase 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.
resultsElectrophoresis 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.
conclusionIn 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.
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