重组人血小板第4因子在大肠杆菌中的表达及其性质分析(英文)  

Expression of Recombinant Human Platelet Factor 4 in Escherichia coli and Its Characterization

作　　者：顾学范 韩芝燕 陆雁 JacquesCaen 韩忠朝 

机构地区：[1]拉里布瓦西埃医院血管和血液研究所 [2]上海贝特生物技术有限公司

出　　处：《中国实验血液学杂志》1996年第2期144-151,共8页Journal of Experimental Hematology

摘　　要：利用PCR和重组DNA技术,在大肠杆菌中得到高效表达的人血小板第4因子(rhPF4)。rhPF4的分离纯化经肝素亲和层析和高效液相层析二步法实现。纯化产物进行了免疫学鉴定、蛋白质银染测定、N-末端氨基酸序列分析和活性分析,结果得到纯度高且具有与天然PF4一样的抑制巨核细胞生成活性的rhPF4。此方法为开发新药和深入研究PF4的作用机制奠定了基础。The polymerase chain reaction (PCR) and recombinant DNA cloning in prokaryotic gene vector were combined to achieve a high expression of recombinant human platelet factor 4 (rhPF4) in Escherichia coli (? coli) . The heparin-sepharose column and a C4 column on reverse phase high per-formance liquid chromatography (RP-HPLC) were used to purify rhPF4 to homogeneity. Purified rhPF4 was biochemically characterized by sodium dodec.vl sulfate (SDS)-polyacrylamide gel electrophoresis (SDS-PAGE), Western blot analysis using specific antibody and N-terminal sequence analysis. Bioassay has shown that the rhPF4, like native PF4, has an inhibitory activity on megakaryocytopoiesis in vitro and in vivo in mice. These data demonstrate that the rhPF4 prepared by our methods is highly purified and biologically active and suggest that a potential use of PF4 as a pharmaceutical agent is available.

关 键 词：血小板第4因子 巨核细胞生成 基因表达 造血前体细胞 

分 类 号：R346[医药卫生—基础医学]