TAT EGFP fusion protein expression and the activity of PC12 cells transduction.docVIP

 PAGE \* MERGEFORMAT 16 TAT EGFP fusion protein expression and the activity of PC12 cells transduction Authors: Hai-Zhen Wang Xu Yu-ming Li Zeng-Fu Yang Jing Yun-Han Zhang 【Abstract】 Objective: To construct pET28a  TAT  EGFP recombinants observed the expression of fusion protein TAT  EGFP in the cell transduction activity. Methods: synthetic encoded TAT protein transduction domain of the DNA fragment into vector pET28a and then connect EGFP gene, the composition of pET28a  TAT  EGFP recombinant child. transformed into E. coli, IPTG-induced TAT  EGFP fusion protein expression, histidine affinity chromatography purification. the fusion protein was added into the culture of PC12 cells into cells observed fluorescence circumstances. RESULTS: The successfully constructed a high expression of pET28a  TAT  EGFP recombinant, purified molecular mass of about 29 ku of fusion protein TAT  EGFP, and cultured PC12 cells confirmed that TAT  EGFP fusion protein through the capacity of biofilm. Conclusion: Through the TAT  EGFP fusion protein expression, purification and activity analysis, confirmed the TAT protein transduction for the peptides and biological macromolecular drugs into tissue cells play a role in the treatment provides a theoretical basis. 【Key Words】 TAT protein transduction domain of recombinant fusion proteins PC12 cells, 0 Introduction With the rapid development of biotechnology, protein and peptide drugs have appeared, but because of the role of biofilm barrier, only a few small molecules to enter the organization, and many have a therapeutic effect of exogenous biological macromolecules to penetrate biological membrane to achieve the effective concentration in the lesion, thus causing difficulties in the treatment of many diseases. In recent years, from the discovery of a human immunodeficiency virus HIV  1 transcription activator (trans  activa  tor, TAT), can effectively guide the peptide or penetrate the cell