Production in Escherichia coli of human thymosin beta 4 as chimeric protein with human tumor necrosis factor |
| |
Authors: | Y Tsuji N Kitahara-Tanabe K Noguchi T Gatanaga D Mizuno G Soma |
| |
Institution: | Biotechnology Research Center, Teikyo University, Kanagawa, Japan. |
| |
Abstract: | We have produced thymosin beta 4 protein in Escherichia coli as a chimeric protein with tumor necrosis factor (TNF). The protein was abundantly expressed, was immunoreactive against both anti-thymosin beta 4 and anti-TNF antibodies, and retained cytotoxicity in a TNF assay using mouse L929 fibroblasts. This latter characteristic enabled the easy and simple purification of thymosin beta 4 merely by following the TNF activity. The chimeric protein was designed to have an Asp-Pro bridge between thymosin beta 4 and TNF which could be specifically cleaved under suitable acidic conditions to release the thymosin beta 4 from the chimeric protein. These results indicate that the expression system in E.coli of a chimeric protein composed of thymosin beta 4 and TNF is appropriate for obtaining an abundant amount of the beta 4 peptide, especially since its purification from tissues is usually difficult because of limited yield and obscurity of its biological activity. |
| |
Keywords: | |
|
|