Expression Strategy of Soluble Recombinant Human TGF-β3 in Escherichia coli: sfGFP -Fusion Tag
نویسندگان
چکیده
Transforming growth factor-beta 3 (TGF-β3) is an important cytokine involved in various biological processes. TGF-β3 used as a scar-reducing antifibrotic agent for acute and chronic wounds fibrosing disorders. TGF-β3, valuable therapeutic protein, produced recombinantly different expression systems. the Escherichia coli (E. coli) system, widely due to its advantages recombinant production, commercially available. However, main problem encountered protein E. cells formation of inclusion body. Various approaches have been developed solve this problem. The use fusion tag one most powerful strategies obtain soluble active form system. Superfolder GFP (sfGFP) tags increase solubility partner coli. In study, with sfGFP (sfGFP-TGFβ3) was successfully BL21 (DE3) high yield purity first time. Purified identified by western blot SDS-PAGE. 20 mg 98% obtained from 1 L bacterial culture. It determined that did not cytotoxic effect on BJ normal human skin fibroblast cells. impact sfGFP-TGFβ3 wound healing evaluated vitro scratch assay. results showed system has potential support process.
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ژورنال
عنوان ژورنال: Sakarya University Journal of Science
سال: 2023
ISSN: ['1301-4048', '2147-835X']
DOI: https://doi.org/10.16984/saufenbilder.1096298