NDLI: Optimization Studies on Prokaryotic Cell Expression of the Human Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL)

Content Provider	Springer Nature Link
Author	Hao, Lin Zhang, Zhi Guo Shi, Zheng Duo Pang, Kun Zhang, Jun Jie Dong, Yang Han, Cong Hui
Copyright Year	2015
Abstract	The aim of the study was to optimize the in vitro induction and expression of the human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and also study the processes of its denaturation, renaturation, and purification. The pGEX-6P-1/TRAIL$_{114–281}$ plasmid was induced by isopropyl-β-d-1-thiogalactopyranoside (IPTG) in Escherichia coli BL21 (DE3), and the expressed target protein was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The protein expressed in the form of inclusion body was first denaturalized and then renaturalized by dilution and dialysis technique. GST-rTRAIL$_{114–281}$ fusion protein was purified by Glutathione-Superflow Resin affinity chromatography and confirmed by Western blot. The molecular weight of GST-rTRAIL expressed in E. coli BL21 (DE3) was approximately 40 kDa. GST-rTRAIL was mainly expressed in the form of inclusion bodies. An optimum expression was induced by IPTG at a concentration of 0.2 mM for 8 h at 37 °C. Glutathione-Superflow Resin affinity chromatography yielded the purified GST-rTRAIL protein which was confirmed by Western blot using anti-GST mouse monoclonal antibody. The optimum prokaryotic cell expression of the human GST-rTRAIL was obtained by 0.2 mM IPTG induction for 8 h at 37 °C. The denatured inclusion body protein can be refolded by dilution and dialysis and purified by Glutathione-Superflow Resin affinity chromatography.
Starting Page	275
Ending Page	279
Page Count	5
File Format	PDF
ISSN	10859195
Journal	Cell Biochemistry and Biophysics
Volume Number	73
Issue Number	2
e-ISSN	15590283
Language	English
Publisher	Springer US
Publisher Date	2015-03-04
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	TRAIL Inclusion body Prokaryotic expression Purification Biochemistry Pharmacology/Toxicology Biotechnology Cell Biology Biological and Medical Physics, Biophysics
Content Type	Text
Resource Type	Article
Subject	Cell Biology Medicine Biochemistry Biophysics

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