Objective To evaluate the construction of expression vector for fusion protein of cell-penetrating pep-tide CCL (PEP-CCL).Methods CCL6-PEP-6XHis was inserted into plasmid pABP,pABP-CCL6-PEP plasmid was extracted and then transfected into HEK293 cells,CCL6-PEP-6XHis was expressed and purified by chromatog-raphy and detected with Western Blot.Results PEP-CCL express vector was successfully constructed and purified. PCR product of CCL6-PEP-6XHis Tag was ligated with T vector,recombinant was transferred into the host cells, then host cells were cultured,plasmid was extracted and sequenced,the sequence was identical to targeted gene. CCL6-PEP-6XHis was successfully inserted into the eukaryotic expression vector pABP,plasmid was extracted and digested,electrophoresis results revealed that a fragment with 430bp was digested by Hind Ⅲ+XbaⅠ,which was identical to the expected value.Western Blot revealed that CCL6-PEP fusion protein could be recognized by His monoclonal antibody.Conclusion PEP-CCL express vector can be constructed and expressed in eukaryotic cells.%目的：评估细胞穿透肽 CCL 融合蛋白构建的可能性。方法将 CCL6-PEP-6XHis 构建至 pABP 质粒，然后提取 pABP-CCL6-PEP 质粒进行人胚肾 HEK293细胞转染表达，以及 CCL6-PEP-6XHis 蛋白层析纯化和检测。结果成功构建并纯化细胞穿透肽 CCL 融合蛋白。将 CCL6-PEP-6XHis Tag 基因经 PCR 扩增、接入 T 载体、克隆、培养，并提取质粒进行测序鉴定，所得序列与目的基因一致。成功将 CCL6-PEP-6XHis 基因构建至哺乳动物细胞表达载体 pABP 中，经质粒提取和酶切鉴定，电泳结果显示，HindⅢ＋ XbaⅠ切出约430 bp 的条带，符合预期，酶切鉴定正确。蛋白质印迹法（Western Blot）检测结果阳性，表明纯化得到的目标蛋白带有 hisx6标签。结论细胞穿透肽 CCL 融合蛋白能够人工构建，并通过真核细胞进行表达。
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