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Cloning and Expression of a Human Tissue Plasminogen Activator Variant: K2s in Escherichia Coli Publisher Pubmed



Dormiani K1 ; Khazaie Y1 ; Sadeghi HMM1, 2 ; Rabbani M1, 2 ; Moazen F2
Authors
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Authors Affiliations
  1. 1. Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran
  2. 2. Department of Pharmaceutical Biotechnology, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran

Source: Pakistan Journal of Biological Sciences Published:2007


Abstract

The DNA sequence of Kringle-2 and serine protease domains of the human tissue plasminogen activator (reteplase, K2S) was PCR amplified. This product was then cloned into the expression vector pET15b plasmid. The presence of the insert was confirmed by restriction digestion, PCR and determination of the nucleotide sequence. By using isopropyl β-D thiogalactopyranoside (IPTG), reteplase was induced in E. coli BL21 cells and analyzed using polyacrylamide gel electrophoresis (PAGE). © 2007 Asian Network for Scientific Information.