PTD—bcr／abl融合癌蛋白片段的表达及其跨膜转运

目的 探讨蛋白转导结构域（PTD）介导的转导bcr/abl癌蛋白片段的方法，为进行免疫治疗提供实验依据。方法 合成编码PTD的基因片段，并与PCR扩增的慢性粒细胞白血病癌蛋bcr/abl基因片段融合，在胸有成竹肠杆菌中表达，将纯化的融合蛋白PTD-bcr/abl与HL-60细胞共孵育，用Western blot分析融合蛋白的跨膜转运。结果 PTD基序可介导bcr/abl蛋白由细胞外跨膜转导进行细胞内。结论 这一结果可能为应用外源蛋白负载（loading)抗原提呈细胞（APC）提供新的途径。

Expression of PTD-bcr/abl fusion oncoprotein fragment carrying protein transduction domain and its transmembrane transportation

Aim To explore the method of transfering oncoprotein bcr/abl, mediated by protein transduction domain (PTD), in order to provide the experimental basis for immune therapy. Methods DNA fragment encoding PTD was synthesized and fused with PCR-amplified bcr/abl gene fragment. The fusion protein was expressed in E.coli as His-tagged protein. The purified fusion protein was loaded to cultured HL-60 cells. Whether the fusion protein entered into HL-60 cells was determined by Western blot. Results It is shown that PTD could mediate transportation of bcr/abl protein to enter into cells. Conclusion These results may provide a new approach for loading exogenous proteins to antigen presenting cells.