摘要
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
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. [
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2003年第3期199-202,共4页
Chinese Journal of Microbiology and Immunology
基金
国家"8 6 3"资助项目 ( 2 0 0 1AA2 15 0 6 1)
