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Volume 14, Issue 2 (5-2011)                   J Arak Uni Med Sci 2011, 14(2): 72-80 | Back to browse issues page

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Moradkhani A, Abtahi H, pakzad I, Karimi M. Antigenic characteristics of recombinant hyaluronidase A of Streptococcus pyogenes expressed in E. coli . J Arak Uni Med Sci. 2011; 14 (2) :72-80
URL: http://jams.arakmu.ac.ir/article-1-783-en.html
1- ARAK AZAD University
2- Arak University of Medical Sciences
3- Ilam Medical Sciences Universiyy , pakzad_i2006@yahoo.com
Abstract:   (14058 Views)
Background: Hyaluronidase A is an antigenic protein that is secreted by Streptococcus pyogenes. Nowadays, streptococcal infections are diagnosed by tracking down anti-hyaluronidase A antibodies. In this study, the attempt was made to generate recombinant hyaluronidase A in E. coli. Materials and Methods: In this experimental study, through designing specific primers and polymerase chain reaction (PCR), hyaluronidase A gene was amplified and after purification, it was sub-cloned in plasmid expression vector pET32a. Then pET32a-hylA was transferred to E. coli BL21-DE3-plySs. Protein generation induced by IPTG. The recombinant protein was purified by Ni-NTA kit and its concentration was assayed by Bradford method. Western-Blot analysis was run for verifying the recombinant hyaluronidase A. Results: The nucleotide sequencing of the gene amplified by PCR was the same as hyaluronidase A gene from Streptococcus pyogenes. Production of the recombinant hyaluronidase A via induction by pET32a-hylA plasmid was done through IPTG. The expressed protein was purified by affinity chromatography by Ni-NTA resin. The concentration of purified protein was 500µg/ml. analysis using a mouse anti-hyaluronidase A serum was reacted with the generated protein using Western-Blot analysis. Conclusion: Recombinant HylA protein can be generated in E.coli and the resulting protein maintains its antigenic properties desirably.
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Subject: Basic Sciences
Received: 2010/07/4

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