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将RT－PCR扩增的人粒细胞集落刺激因子（hG－CSF）结构基因与表达载体pJGW1重组，转化含有pGP1－2质粒的E.coliDH5α，进行温度诱导表达。经SDS－PAGE分析，rhG－CSF表达量占菌体总蛋白量的30％以上。将其复性，经CM－52FF离子交换层析纯化，纯化后的rhG－CSF（纯度＞98％）经SDS－PAGE测定分子量约为19kD；经胰酶裂解、反相HPLC分析肽谱具有8条特异肽段；等电聚焦测定PI值约为5．8；免疫印迹实验证实其与标准hG－CSF具有相同的抗原反应特异性；NH2-末端氨基酸分析与文献报道一致，采用小鼠白血病细胞株NFS－60测定活性为I×108IU／mg。
The hG-CSF structural gene amplified by RT-PCR was recombined with the expression vector pJGW1 and transformed into E. coli DH5α containing pGP1-2 plasmid for temperature-induced expression. The SDS-PAGE analysis showed that rhG-CSF accounted for more than 30% of the total bacterial protein. The recombinant protein was purified by CM-52FF ion-exchange chromatography. The purified rhG-CSF (purity> 98%) had a molecular weight of about 19kD by SDS-PAGE. The peptide was identified by reverse phase HPLC The specific PI was about 5.8. The immunoprecipitation assay confirmed that it had the same antigen specificity as the standard hG-CSF. The NH2-terminal amino acid analysis was consistent with that reported in the literature. Using mouse leukemia cells The activity of strain NFS-60 was determined to be Ix 108 IU / mg.