ANTIGENIC CHARACTERISTICS OF RECOMBINANT HYALURONIDASE A OF STREPTOCOCCUS PYOGENES EXPRESSED IN E. COLI

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MORADKHANI A.; ABTAHI H.; PAKZAD I.; KARIMI MASOUDEH

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Journal: Journal of Arak University Medical Sciences Year:2011 | Volume:14 | Issue:2 (55) Page(s): 72-80

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Title

ANTIGENIC CHARACTERISTICS OF RECOMBINANT HYALURONIDASE A OF STREPTOCOCCUS PYOGENES EXPRESSED IN E. COLI

Author(s)

MORADKHANI A. | ABTAHI H. | PAKZAD I. | KARIMI MASOUDEH | Issue Writer Certificate

Keywords

E. COLIQ2

GENE EXPRESSIONQ2

HYALURONIDASE A GENEQ2

STREPTOCOCCUS PYOGENESQ2

Abstract

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 500mg/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.

Cites

PRODUCTION AND PURIFICATION OF ENZYMATIC REGION OF STREPTOCOCCAL HYALURONIDASE IN E.COLI

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APA: Copy

MORADKHANI, A., ABTAHI, H., PAKZAD, I., & KARIMI, MASOUDEH. (2011). ANTIGENIC CHARACTERISTICS OF RECOMBINANT HYALURONIDASE A OF STREPTOCOCCUS PYOGENES EXPRESSED IN E. COLI. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), 14(2 (55)), 72-80. SID. https://sid.ir/paper/68989/en

Vancouver: Copy

MORADKHANI A., ABTAHI H., PAKZAD I., KARIMI MASOUDEH. ANTIGENIC CHARACTERISTICS OF RECOMBINANT HYALURONIDASE A OF STREPTOCOCCUS PYOGENES EXPRESSED IN E. COLI. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ)[Internet]. 2011;14(2 (55)):72-80. Available from: https://sid.ir/paper/68989/en

IEEE: Copy

A. MORADKHANI, H. ABTAHI, I. PAKZAD, and MASOUDEH KARIMI, “ANTIGENIC CHARACTERISTICS OF RECOMBINANT HYALURONIDASE A OF STREPTOCOCCUS PYOGENES EXPRESSED IN E. COLI,” ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), vol. 14, no. 2 (55), pp. 72–80, 2011, [Online]. Available: https://sid.ir/paper/68989/en

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