带有蛋白转导结构域的Bcr/Abl癌蛋白片段的表达及其跨膜转运

HIV 1编码的反式激活蛋白TAT具有将细胞外蛋白转导进入细胞的基序 ,称为蛋白转导结构域 (PTD) .为研究PTD介导的PTD Bcr Abl融合蛋白的跨膜转运 ,合成了编码PTD的基因片段 ,并与PCR扩增的慢性粒细胞白血病癌蛋白bcr abl基因片段融合 .在大肠杆菌中表达纯化了融合蛋白 ,将纯化的融合蛋白加入培养的HL60细胞和C2C12细胞后 ,发现PTD基序可以介导Bcr Abl蛋白自由从细胞外跨膜转导进入细胞内 .研究结果可能为用外源蛋白负载 (loading)免疫活性细胞如抗原提呈细胞提供新的途径 .

Expression of Bcr/Abl Oncoprotein Fragment Carrying Protein Transduction Domain and Its Transportation through the Plasma Membrane

Trans activating protein TAT encoded by HIV\|1 contains a motif called protein transduction domain (PTD),which mediates transportation of extra\|cellular proteins into cytoplasm.DNA fragment encoding PTD was synthesized and fused to PCR\|amplified bcr/abl gene fragment, which encode an onco\|protein critical to chronic myeloid leukemia.The fusion peptide was expressed in E.coli as His\|tagged protein.The purified fusion protein was loaded to cultured HL60 cells and C2C12 cells.Using Western blot and immunocytochemistry techniques,it was shown that PTD could mediate transportation of Bcr/Abl protein into cells.These results may provide anew approach of loading exogenous proteins to immuno\|active cells such as antigen presenting cells.