title

کلون، بیان، خالص سازی و بهینه سازی بیان آنزیم آلفا-انولاز از باکتری استافیلوکوکوس اورئوس در ایشرشیاکولای

قاسمی, یونس and حاجی قهرمانی, نسیم and دباغ, فاطمه and قشون, محمد باقر and یاراحمدی, الهام and مباشر, محمد علی and منتظری نجف آبادی, نیما (1394) کلون، بیان، خالص سازی و بهینه سازی بیان آنزیم آلفا-انولاز از باکتری استافیلوکوکوس اورئوس در ایشرشیاکولای. Minerva Biotechnologica ــ 28 (1). pp. 33-38. شاپا 11204826

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Title

Cloning, expression, purification and expression condition optimization of α-enolase from Staphylococcus aureus in Escherichia coli

English Abstract

The multifunctional enzyme α-enolase belongs to a family of cytoplasmic and glycolytic enzymes, which is localized in the cytoplasm and at the surface of many eukaryotic and prokaryotic cells. α-enolase on the surface of Staphylococcus aureus is a receptor with high affinity for laminin, the most abundant extracellular matrix component. Laminin-binding ability plays a considerable role in the pathogenesis of this microorganism. S. aureus is an opportunistic pathogen that causes major nosocomial infections and variety of diseases in human beings. This organism has a strong tendency to develop antibiotic resistance. Its property to spread of antibiotic resistance has intensified the need of novel antistaphylococcal techniques. α-enolase as an important surface antigen, is a potential vaccine or immunotherapeutic candidate. For cloning and expression studies, α-enolase gene was amplified by PCR using the specific primers. Then it was inserted into the pET-15b vector and transformed in to Escherichia coli DE3. Gene expression was induced at 30°C using IPTG and the recombinant enzyme purification carried out by Ni-NTA Spin Columns. Protein was shown with SDS-PAGE analysis and then, different induction conditions for optimization of the recombinant protein expression were used. The highest protein expression was observed when the α-enolase production was induced using the mixture of two different inducers at the same time.

Item Type:Article
زبان سند : انگلیسی
نویسنده مسئول :یونس قاسمی
نویسنده :نسیم حاجی قهرمانی
نویسنده :فاطمه دباغ
نویسنده :محمد باقر قشون
نویسنده :الهام یاراحمدی
نویسنده :محمد علی مباشر
نویسنده :نیما منتظری نجف آبادی
Additional Information:Impact factor: 0.25 Indexed in: Science Citation Index Expanded (ISI), Scopus, EMBASE,
کلیدواژه ها (انگلیسی):Gene cloning, α-enolase, Vaccine candidate, Staphylococcus aureus, Escherichia coli
Subjects:QV pharmacology > QV 737 Pharmaceutical Biotechnology
Divisions:School of Pharmacy > Department of Pharmaceutical Biotechnology
ID Code:12444
Deposited By: دکتر نسیم حاج قهرمانی
Deposited On:19 Dec 1398 10:21
Last Modified:19 Dec 1398 10:21

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