Prepared two ways with the original expression TAT protein transduction domain fusion protein compared penetrating ability.docVIP

 PAGE \* MERGEFORMAT 20 Prepared two ways with the original expression TAT protein transduction domain fusion protein compared penetrating ability Of: Xu Xun, Lin Xin, Tian Hong, Zhao Qian-Qian, Ji Bao Ju, Liu Jing, Xia Sheng, Wang Shengjun, Xu of the river, Shaoqi Xiang [Abstract] Objective: To evaluate a variety of techniques combined with different ways of preparation of TAT transduction domain eGFP transmembrane protein subcellular localization efficiency and to establish an effective, stable and efficient preparation of transmembrane proteins. Methods: induced , purified fusion protein inclusion bodies were direct renaturation desalination and dialysis, and with the cell co-culture, and then by flow cytometry, confocal microscopy and Western blot evaluation of penetrating ability of the fusion protein. Results: dialysis Preparation of the fusion protein of penetrating efficiency is very low, the purified fusion protein directly after desalting by flow cytometry, confocal microscopy and Western blot analysis and other methods that have a high penetrating ability. Conclusion: The preparation of two ways membrane fusion protein can wear, but the dialysis of the preparation of the transmembrane protein, low efficiency, and not easily localized in the nucleus, the impact of further protein in the nucleus of the biological behavior, and desalination of denatured proteins with high efficiency transmembrane efficiency. [Keywords:] transduction domain, dialysis, refolding, inclusion bodies, penetrating efficiency [Abstract] Objective: To evaluate the efficacy and subcellular localization of the fusion protein eGFP interest protein including protein transduction domain of TAT by multi technology and to obtain the more effective cellular uptake protein for the further research of its biological function.Methods: Induced the Expression of fusion protein, and extracted and purified it from the inclusion bodies. Jurkat cells exposure to fusion protein which