原核表达的含PTD-eGFP不同分子质量融合蛋白穿膜能力的比较

Comparison the transduction ability of three prokaryotic expression of fusion protein which contains of protein transduction domain and eGFP

目的:评价HIV-1 TAT蛋白转导结构域(PTD)携带的不同分子质量融合蛋白的穿细胞膜能力。方法:诱导表达、纯化含PTD-eGFP的不同分子质量的融合蛋白,与细胞共育后,通过流式细胞术、激光共聚焦评价融合蛋白穿膜能力。结果:诱导表达、纯化获得3种含PTD-eGFP的不同分子质量的融合蛋白。流式细胞术和激光共聚焦方法分析发现3种融合蛋白均具有高效穿膜能力。结论:PTD介导融合蛋白的穿膜能力与分子质量无直接关系。

Objective： To judge the efficacy of the fusion protein including protein transduction domain of HIV TAT by multi-biotechnology and to obtain the more effective cellular uptake protein for the further research of its bio-funetion. Methods： The expression of three fusion proteins were induced by IPTG and were purified from the inclusion bodies. Flow cytometry analysis and confocal microscope observation were used to evaluate the efficacy of the fusion proteins to transduce into the Jurkat cells, when the cells were exposed to fusion protein. Results： The fusion proteins could transduce into Jurkat T cells efficiently. Conclusion： All the three fusion proteins could cross the cell membrane and had no difference.