Paper Information

Journal:   ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ)   APRIL-MAY 2011 , Volume 14 , Number 1 (54); Page(s) 93 To 103.
 
Paper: 

THE EXPRESSION OF RECOMBINANT STREPTODORNASE IN E.COLI BACTERIA

 
 
Author(s):  KAMANI M., ABTAHI HAMID*, MOSAYEBI GHASEM, NAZARI R., KARIMI MASOUDEH
 
* MEDICAL AND MOLECULAR RESEARCH CENTER, ARAK UNIVERSITY OF MEDICAL SCIENCES, ARAK, IRAN
 
Abstract: 

Background: In pyoderma infections, the density of pus is related to desoxiribonucleoproteins. The use of streptodornase (DNase) in combination with streptokinase can help dissolve purulent secretions of infections which results in healing the wound through the discharge of pus from the necrotic tissue. The aim of this study was to produce recombinant streptodornase from group A strain of Streptococcus pyogenes which is highly efficient in terms of active streptodornase production using expression vector.
Materials and Methods: In this applied-fundamental study, genomic DNA of streptodornase gene (sd) was extracted by phenol-chloroform. Then by using specific primers of streptodornase gene, it was amplified through PCR. The resulting streptodornase gene was cloned in pGEX4T1-sd transformer for expression and the pGEX4T1-sd plasmid was transferred to the sd. E.coli BL21. Protein production was done by induction via IPTG and optimization of the conditions. The recombinant protein was purified using the glutathione sepharose 4B kit.
Results: The nucleotide sequence of PCR and group A streptodornase Streptococcus was totally the same. The production of the streptodornase recombinant protein was done by inducing pGEX4T1-sd plasmid via Isopropyl
b-D-1-thiogalactopyranoside. Protein purification was done through affinity-chromatography by using glutathione sepharose 4B. The recombinant protein was reacted with anti-streptodornase mouse serum through Western-Blot method.
Conclusion: Recombinant streptodornase can be produced by pGEX4T1 in E. coli. The recombinant protein maintains its antigenic property desirably. Noticing the domestic need in Iran, low rate of production, and pathogenesis of streptococci, production of this recombinant product is feasible.

 
Keyword(s): GENE EXPRESSION, STREPTOCOCCUS GROUP A, STREPTODORNASE
 
References: 
  • ندارد
 
  Yearly Visit 29
 
Latest on Blog
Enter SID Blog