1. [Prokaryotic expression and biological activity of recombinant human IL-17F/His protein].
- Author
-
Liu GQ, Wu HY, Li LB, Ma ZN, Lu PR, and Zhang XG
- Subjects
- Blotting, Western methods, Cell Proliferation, Cells, Cultured, Cloning, Molecular methods, HeLa Cells metabolism, HeLa Cells pathology, Humans, Interleukin-17 chemistry, Interleukin-17 genetics, Monocytes, Recombinant Proteins genetics, Recombinant Proteins isolation & purification, Recombinant Proteins metabolism, Cytokines metabolism, Interleukin-17 isolation & purification, Interleukin-17 metabolism, Interleukin-6 metabolism, Tumor Necrosis Factor-alpha metabolism
- Abstract
Aim: To Prepare recombinant human IL-17F/His protein and investigate its biological activity in vitro., Methods: The gene region of human IL-17F was cloned by RT-PCR. After identification by sequencing, the hIL-17F gene encoding function domain was cloned into expression plasmid PQE3.0 and transfected into E.coli M15. By the induction of Isopropyl-β-D-Thiogalacto-Pyranoside(IPTG), recombinant IL-17F/His protein was effectively expressed in E.coli M15. The recombinant protein was identified by Western blot., Results: After renaturation and purification by HiTrap(TM); affinity column, the recombinant protein can up-regulate macrophages to secret TNF-α, IL-6 and other relative cytokines. It also promoted proliferation of HeLa cells in vitro., Conclusion: hIL-17F/His recombinant protein is of high biological activity, which can be used to make further study of its special characteristic.
- Published
- 2011