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Cloning, expression, and cost effective purification of authentic human epidermal growth factor with high activity
المؤلفون المشاركون
Pouranvari, Sarah
Ibrahimi, Fayruz
Maddah, Bozorgmehr
Javadi, Gholamreza
المصدر
Iranian Red Crescent Medical Journal
العدد
المجلد 18، العدد 3 (31 مارس/آذار 2016)، ص ص. 1-8، 8ص.
الناشر
تاريخ النشر
2016-03-31
دولة النشر
الإمارات العربية المتحدة
عدد الصفحات
8
التخصصات الرئيسية
الملخص EN
Background : Epidermal growth factor (EGF) plays a fundamental role in the healing of wounds relating to skin damage, the cornea, and the gastrointestinal tract.
Objectives : The aim of this study is the cloning, expression, and purification of recombinant human EGF (rhEGF), and an assessment of its activity.
Materials and Methods: In the present experimental study, a synthetic pET28a (+) -hEGF construct was prepared.
In order to ligate hEGF into pET24a (+), the PCR technique was performed, using special primers that possess restriction enzyme sites, which are also located in appropriate sites in pET24a (+).
After transferring this construct into E.
coli cells, protein expression was performed under standard conditions.
Protein solubilization was done by urea.
hEGF purification and refolding were carried out using gradient dialysis against the urea.
We used RP-HPLC to compare between rhEGF and commercial rhEGF as a control.
Finally, an MTT assay was performed to assess the viability of the NIH 3T3 cells treated with various concentrations of rhEGF.
Results : Dialysis after urea solubilization caused precipitation of unwanted proteins, resulting in achievement of purified EGF with > 90% purity, without the need for expensive and time-consuming process.
The MTT assay results showed that our rhEGF activate significantly higher proliferation of NIH 3T3 cells in comparison to the control (P-values were < 0.0001), in total concentrations and times evaluated Conclusions: Via our purification protocol, a sufficient amount of bioactive recombinant human epidermal growth factor was obtained in just a few affordable steps, with superlative purity.
نمط استشهاد جمعية علماء النفس الأمريكية (APA)
Pouranvari, Sarah& Ibrahimi, Fayruz& Javadi, Gholamreza& Maddah, Bozorgmehr. 2016. Cloning, expression, and cost effective purification of authentic human epidermal growth factor with high activity. Iranian Red Crescent Medical Journal،Vol. 18, no. 3, pp.1-8.
https://search.emarefa.net/detail/BIM-679286
نمط استشهاد الجمعية الأمريكية للغات الحديثة (MLA)
Pouranvari, Sarah…[et al.]. Cloning, expression, and cost effective purification of authentic human epidermal growth factor with high activity. Iranian Red Crescent Medical Journal Vol. 18, no. 3 (Mar. 2016), pp.1-8.
https://search.emarefa.net/detail/BIM-679286
نمط استشهاد الجمعية الطبية الأمريكية (AMA)
Pouranvari, Sarah& Ibrahimi, Fayruz& Javadi, Gholamreza& Maddah, Bozorgmehr. Cloning, expression, and cost effective purification of authentic human epidermal growth factor with high activity. Iranian Red Crescent Medical Journal. 2016. Vol. 18, no. 3, pp.1-8.
https://search.emarefa.net/detail/BIM-679286
نوع البيانات
مقالات
لغة النص
الإنجليزية
الملاحظات
Includes bibliographical references : p. 8
رقم السجل
BIM-679286
قاعدة معامل التأثير والاستشهادات المرجعية العربي "ارسيف Arcif"
أضخم قاعدة بيانات عربية للاستشهادات المرجعية للمجلات العلمية المحكمة الصادرة في العالم العربي
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