人类新细胞因子(CKLF-1)在大肠杆菌中的表达与鉴定

目的：进一步研究人类新细胞因子－－趋化素样因子1（CKLF－1）蛋白的结构与生物学活性。方法：构建了融合蛋白的原核表达质粒，并在大肠杆菌中进行诱导表达，获得了原核表达的融合蛋白，经凝血酶切割获非融合重组蛋白，并进行体外活性鉴定。结果：CKLF－1原核表达蛋白获得了高表达，纯化后获得约90％纯度以上蛋白，发现其可以与肝素结合，生物学活性检测表明其对人中性粒细胞，外周血单个核细胞和大鼠的巨噬细胞具有明显的趋化作用。结论：CKLF－1原核表达蛋白具有与真核表达蛋白类似的生物学活性。

Expression and identification of a novel human cytokine CKLF-1 in E.coli

Objective To further study the structural and functional characteristics of a novel human cytokine, chemokine-like factor 1 (CKLF-1). Method By using recombinant DNA technique, the recombinant MS2-CKLF-1 fusion protein containing thrombin linker was expressed, digested with thrombin, and purified with heparin affinity chromatography. The Boyden chamber was used for the bioassay of recombinant CKLF-1. Result The recombinant CKLF-1 expressed in E.coli with high efficiency. Up to 90% purity was reached by heparin affinity chromatography. The CKLF-1 protein exhibits significantly chemotactic effect on PBMC, U937 cells and neutrophils of human and macrophages of rat. Conclusion The recombinant CKLF-1 from E.coli has similar bioactivity as that from eukaryocytic expression system. [