Construction and in vitro activity evaluation of protein transduction domain-transactivator of transcription and Candida antarctica lipase B fusion proteins

Rosetta (DE3) cells to express the labeled protein calB-Tat. Protein concentrations were measured using a commercial BCA kit, and the transmembrane activity of the proteins in SH-SY5Y cells was observed under a fluorescence-inverted microscope. MTT and Western blotting assays were conducted to examine toxicity. The fusion protein exhibited low toxicity. As the concentration of the fusion protein decreased, the effect on cell viability decreased. Additionally, the fusion protein penetrated the cell membrane penetration was stable and was specifically expressed in cells. Taken together, the pET28a-calB-Tat prokaryotic vector was generated, yielding a significant amount of the calB-Tat protein. This increased the cell membrane and perhaps reveals a new way of delivering weight-loss drugs and protein-based medications.