title: Cloning, Expression, and Purification of Recombinant Lysostaphin From Staphylococcus simulans.
abstract: BACKGROUND
  Staphylococcus aureus is one of the most common causes of nosocomial infections and its resistance to antibiotics is a global concern. Lysostaphin is an antimicrobial agent belonging to a major class of antimicrobial peptides and proteins known as the bacteriocins. It exhibits a high degree of anti-staphylococcal bacteriolytic activity.
  
  
  OBJECTIVE
  In this study, high level of recombinant mature lysostaphin in Escherichia coli was produced by using pET32a expression vector.
  
  
  METHODS
  The S. simulans gene encoding lysostaphin was extracted, amplified by polymerase chain reaction (PCR), and sub-cloned in prokaryotic expression vector pET32a. E. coli BL21 (DE3) plysS were transformed with pET32a-lys and gene expression was induced by IPTG. The expressed protein was purified by affinity-chromatography using (Ni-NTA) resin.
  
  
  RESULTS
  PCR and sequencing results confirmed the successful cloning of the target gene into the vector. The expression of protein was induced by IPTG and high concentration of the recombinant protein was obtained via the purification process by affinity-chromatography.
  
  
  CONCLUSIONS
  Our data showed that the recombinant mature lysostaphin protein produced by pET32a vector in E. coli system was very efficient.
authors.affiliation: Department of Biotechnology, Arak University of Medical Sciences, Arak, IR Iran.
authors.forename: Leila
authors.initials: L
authors.lastname: Farhangnia
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors.forename: Ehsanollah
authors.initials: E
authors.lastname: Ghaznavi-Rad
authors.affiliation: Department of Biotechnology, School of Medicine, University of Medical Sciences, Arak, IR Iran.
authors.forename: Neda
authors.initials: N
authors.lastname: Mollaee
authors.affiliation: Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, IR Iran.
authors.forename: Hamid
authors.initials: H
authors.lastname: Abtahi
keywords: Escherichia coli; Lysostaphin; Recombinant Proteins; Staphylococcus