کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5516156 1542308 2017 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Effective non-denaturing purification method for improving the solubility of recombinant actin-binding proteins produced by bacterial expression
ترجمه فارسی عنوان
روش تصفیه بدون انتیاتورین موثر برای بهبود حلالیت پروتئین های اتصال دهنده اکتین نوترکیب تولید شده توسط بیان باکتری
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
چکیده انگلیسی


- The expression and purification condition for truncated actin binding proteins was optimized from Escherichia coli strain.
- The treatment of sarkosyl-detergent increases the solubility of bacterial recombinant proteins.
- Minimizing concentration of sarkosyl was suggested for enhancing the production of soluble proteins.

Bacterial expression is commonly used to produce recombinant and truncated mutant eukaryotic proteins. However, heterologous protein expression may render synthesized proteins insoluble. The conventional method used to express a poorly soluble protein, which involves denaturation and refolding, is time-consuming and inefficient. There are several non-denaturing approaches that can increase the solubility of recombinant proteins that include using different bacterial cell strains, altering the time of induction, lowering the incubation temperature, and employing different detergents for purification. In this study, we compared several non-denaturing protocols to express and purify two insoluble 34 kDa actin-bundling protein mutants. The solubility of the mutant proteins was not affected by any of the approaches except for treatment with the detergent sarkosyl. These results indicate that sarkosyl can effectively improve the solubility of insoluble proteins during bacterial expression.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 133, May 2017, Pages 193-198
نویسندگان
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