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Iranian Journal of Basic Medical Sciences، جلد ۱۷، شماره ۹، صفحات ۶۶۷-۶۷۲
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عنوان انگلیسی Cloning and expression of the enzymatic region of Streptococcal hyaluronidase
چکیده انگلیسی مقاله Objective(s): Streptococcus pyogenes produces extracellular hyaluronidase enzyme. This enzyme is directly associated with the spread of the organism during infection. The objective of the present study was to clone and express the nucleotide sequence of the enzyme which is involved in hyaluronidase enzymatic activity. Materials and Methods: The enzymatic region of hyaluronidase gene was detected by bioinformatics method. The PCR method was used to amplify enzymatic region of hyaluronidase gene from chromosomal DNA of Streptococcus pyogenes. The eluted product was cloned into the prokaryotic expression vector pET32a which was digested by BamHI and HindIII restriction endonuclease enzymes. The target protein was expressed in the Escherichia coli. The bacteria including pET32a-hylA (hylA is abbreviation of Streptococcus pyogenes hyaluronidase gene and hylA is abbreviation of Streptococcus pyogenes hyaluronidase protein) plasmids were induced by IPTG and analyzed by SDS-PAGE. The enzymatic evaluation and antigenicity was finally studied. Results: Enzymes digestion analysis, sequencing results showed that the target gene (1296 base pair) was inserted correctly into the recombinant vector. The expressed protein (65 KDa) was purified successfully via affinity chromatography. Data also indicated that enzymatic region of hyaluronidase protein from Streptococcus pyogenes was recognized in all 5 patient’s sera. Conclusion: In general, it is possible to produce the enzymatic regions of the Streptococcus pyogenes hyaluronidase in E. 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.
کلیدواژه‌های انگلیسی مقاله Gene expression, Hemolytic streptococci, Hyaluronidase
نویسندگان مقاله نفیسه al سادات میرجمالی | nafiseh al sadat mirjamali
department of microbiology, science and research branch, islamic azad university, arak branch, arak, iran

سازمان اصلی تایید شده: دانشگاه آزاد اسلامی علوم و تحقیقات (Islamic azad university science and research branch)

صفیه صوفیان | safieh soufian
biology department, payame noor university, arak, iran

سازمان اصلی تایید شده: دانشگاه پیام نور تهران (Payame noor university)

ندا مولایی | neda molaee
molecular and medicine research center, department of microbiology, school of medicine, arak university of medical sciences, arak, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اراک (Arak university of medical sciences)

شبنم صدوق عباسیان | shabnam sadoogh abbasian
molecular and medicine research center, department of microbiology, school of medicine, arak university of medical sciences, arak, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اراک (Arak university of medical sciences)

حمید ابطحی | hamid abtahi
molecular and medicine research center, department of microbiology, school of medicine, arak university of medical sciences, arak, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اراک (Arak university of medical sciences)

نشانی اینترنتی http://ijbms.mums.ac.ir/article_3329.html
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