Solubility Comparing of Non-Classical Inclusion Bodies in Differ-ent Solubilization Buffers
سال انتشار: 1395
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 586
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شناسه ملی سند علمی:
CBGCONF03_150
تاریخ نمایه سازی: 8 آبان 1395
چکیده مقاله:
There are different non soluble proteins. One solubilization method is not suitable for all of proteins. The reason of our study was to compare different solubilization buffers for solubilizing the inclusion bodies were produced in recombinant E. coli BL21 and choosing the best of them. For this purpose, six different buffers were con-structed and specific amount of inclusion bodies were solubilized in all of them. Solutions were centrifuged then and protein concentration of each supernatant was measured with microBradford assay. The result showed that Solubilization buffer D (50 mM Tris–HCl, 5 % glycerol, 0.1 mM EDTA, 50 mM NaCl, and 0.4 % sarkosyl, β-mercaptoethanol 15 mM pH 7.9) is the best buffer for solubilizing of our non-classical inclusion body. Optimi-zation of solubilization buffer for specific inclusion bodies is very important in purification process of recombi-nant protein produced in E.coli.
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نویسندگان
Sepideh Abbaszadeh
Iranian research organization for science and technology (IROST), biotechnology department, Tehran, Iran
Nahid Bakhtiari
Iranian research organization for science and technology (IROST), biotechnology department, Tehran, Iran
Zahra Amini bayat
Iranian research organization for science and technology (IROST), biotechnology department, Tehran, Iran