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

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MIRJAMALI MEHRABADI N.A.; SOUFIAN S.; ABTAHI H.

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Journal Paper

Paper Information

Journal: Journal of Arak University Medical Sciences Year:2014 | Volume:17 | Issue:3 (85) Page(s): 67-75

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Title

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

Author(s)

MIRJAMALI MEHRABADI N.A. | SOUFIAN S. | ABTAHI H. | Issue Writer Certificate

Keywords

CLONINGQ2

GENE EXPRESSIONQ2

HYALURONIDASEQ2

STREPTOCOCCUS PYOGENESQ2

Abstract

Background: STREPTOCOCCUS PYOGENESproduce extracellular HYALURONIDASE enzyme which is directly associated with the spreading of the organism during infection. HYALURONIDASE enzyme is able to break hyaluronic acid or interstitial cement. This enzyme might be used in cancer treatment.The objective of the present study was to clone and express the nucleotide sequence of this enzyme which is involved in HYALURONIDASE enzymatic activity.Materials and Methods: The enzymatic region of HYALURONIDASE gene was detected by bioinformatics methods. The polymerase chain reaction method was used to amplify the region. The amplified product was cloned into the expression vectorpET32a. E. coli BL21 (DE3) pLYsS was transformed with recombinant plasmids. Then GENE EXPRESSION was induced by IPTG. The expressed protein was purified successfully via affinity chromatography by NiNTA kit. The integrity of the product was confirmed by western-blot analysis.Results: The nucleotide sequence of amplified gene was consistent with the streptocuccal HYALURONIDASE gene. The concentration of recombinant protein calculated to 500 mg purified protein per liter. The enzymatic region of recombinant protein from STREPTOCOCCUS PYOGENES was recognized by all five patient’s sera with Streptococcus infection.Conclusion: In general, it is possible to produce the enzymatic regions of the STREPTOCOCCUS PYOGENEShyaluronidase in Escherichia coli. The antigenic property of the produced protein is well retained. Considering the product's domestic demand and also low efficiency of production and pathogenicity of Streptococcus species, it is possible to produce it as recombinant product.

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APA: Copy

MIRJAMALI MEHRABADI, N.A., SOUFIAN, S., & ABTAHI, H.. (2014). PRODUCTION AND PURIFICATION OF ENZYMATIC REGION OF STREPTOCOCCAL HYALURONIDASE IN E.COLI. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), 17(3 (85)), 67-75. SID. https://sid.ir/paper/68907/en

Vancouver: Copy

MIRJAMALI MEHRABADI N.A., SOUFIAN S., ABTAHI H.. PRODUCTION AND PURIFICATION OF ENZYMATIC REGION OF STREPTOCOCCAL HYALURONIDASE IN E.COLI. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ)[Internet]. 2014;17(3 (85)):67-75. Available from: https://sid.ir/paper/68907/en

IEEE: Copy

N.A. MIRJAMALI MEHRABADI, S. SOUFIAN, and H. ABTAHI, “PRODUCTION AND PURIFICATION OF ENZYMATIC REGION OF STREPTOCOCCAL HYALURONIDASE IN E.COLI,” ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), vol. 17, no. 3 (85), pp. 67–75, 2014, [Online]. Available: https://sid.ir/paper/68907/en

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