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Cloning, expression and purification of recombinant prolysostaphin protein and evaluating its in vitro antistaphylococcal activity
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Leila Farhang nia   , Ehsanollah Ghaznavi Rad , Neda Molaee , Hamid Abtahi |
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Abstract: (7086 Views) |
| Introduction:Staphylococcusaureus causes a wide range of infections, from
skin infections disseminated to systemic
infections leading to organ failure and death. Drug resistance in this group of
pathogens is a world-wide concern and needs the development of novel agents.
Lysostaphin, an example of such a novel agents,is a bacteriocin secreted by Staphylococcussimulans
to kill Staphylococcus aureus through proteolysis of the Staphylococcus
cell wall.The aim of this study was to high level expression and in
vitro evaluation of antistaphylococcal activity of recombinant
prolysostaphinprotein under invitro conditions. Materials and Methods:In this experimental study, lysostaphin gene of Staphylococcussimulans was
amplified by PCR method, then was cloned and expressed into the expression
vector pET-32a. The expressed protein was purified by affinity- chromatography
using (Ni-NTA) resin kit. The Study of antibacterial activity against a cell
suspension of S. aureuswere was performed using turbidity assay. Results: PCR and sequencing results showed the successful cloning
of the target gene into the recombinant vector. The expression of protein was
induced by IPTG and high concentration of the recombinant protein with
antistaphylococcal activitywas purified using Ni-NTA resin. Conclusion:Our data showed that, the pET32a expression vector is a very efficient
system for producing recombinant prolysostaphin protein in E.coli. |
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| Keywords: Staphylococcus aureus, Lysostaphin, Molecular cloning |
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Full-Text [PDF 521 kb]
(1910 Downloads)
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Type of Study: Research |
Subject:
General
Received: 2013/07/16 | Accepted: 2013/12/17 | Published: 2014/06/11
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