کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8360684 | 1542346 | 2014 | 5 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Expression and purification of soluble human APRIL in Escherichia coli using ELP-SUMO tag
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موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
APRIL is a member of the tumor necrosis factor (TNF) family of ligands that mediate tumor cells proliferation as well as survival, depending on the cellular context. In this report, we present a novel method to obtain soluble human APRIL in Escherichia coli using the elastin-like polypeptide and SUMO (ELP-SUMO) tags. The fusion protein with ELP-SUMO tag was expressed in a soluble form at 15 °C. After purification based on inverse transition cycling (ITC) method, the purified ELP-SUMO-hAPRIL fusion protein was subsequently cleaved by SUMO protease to release mature hAPRIL. Following affinity chromatography, the target protein was re-purified with high purity. Finally, about 4.8 mg recombinant hAPRIL was obtained from 1 l bacterial culture with no less than 85% purity. The molecular mass (Mr) of the recombinant hAPRIL was confirmed by MALDI-TOF MS as Mr 16,314. The purified hAPRIL exhibits biological activity on Jurkat cells. It is the first report on soluble production of hAPRIL in E. coli using ELP-SUMO tag.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 95, March 2014, Pages 177-181
Journal: Protein Expression and Purification - Volume 95, March 2014, Pages 177-181
نویسندگان
Jie Zhang, Lei Ma, Shuang Quan Zhang,