PTD-hEGF Fusion Protein—Gene Expression and Function Analysis

Abstract:

To produce membrane-permeable human epidermal growth factor (hEGF)，a pPTD-hEGF prokaryocyte expression vector was constructed and transformed into E. coli BL 21 (DE3). The PTD-hEGF fusion protein was induced by 0.3mmol/L of IPTG expressed in the form of inclusion body with an yield of 40% of the total protein in the cells，and then purified by Ni²⁺-NTA affinity chromatography. The SDS-PAGE analysis showed a single fusion protein band with a molecular weight of 16kD. The amino acid sequence was checked by MALDI-TOF-MS analysis. The genetic engineering PTD-hEGF fusion protein obviously promoted the proliferation and growth of the HEK-293 cells in vitro.

Key words:PTD-hEGF fusion protein，prokaryocyte expression，HEK-293 cell