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A preliminary study on the expression and biological function of recombinant human SCF-TPO fusion protein
Author(s): 
Pages: 19-22
Year: Issue:  1
Journal: CHINESE JOURNAL OF HEMATOLOGY

Keyword:  干细胞因子血小板生成素融合蛋白质类SCF-TPO生物学活性;
Abstract: 目的研究重组人干细胞因子-血小板生成素(SCF-TPO)融合蛋白表达的最佳条件及其生物学活性.方法设计引物,利用RT-PCR从胎肝细胞中扩增到SCF和TPO氨基端功能区片段,采用基因融合新策略将其融合成SCF-TPO融合基因,克隆到pGEM-T载体中,构建pET32a/SCF-TPO融合基因原核表达载体,在宿主菌E.coliBL21(DE3)plysS中经异丙基-β-D-硫代半乳糖(IPTG)诱导其高表达SCF-TPO,SDS-聚丙烯酰胺凝胶电泳和Western blot检测.目的蛋白经包涵体变性,金属螯合层析纯化,透析复性,用细胞因子依赖细胞系MO7e进行细胞增殖实验.结果获得SCF-TPO融合基因的高表达,表达量占菌体总蛋白的30%以上.Western blot法鉴定表达正确,MTT法证明该融合蛋白具有细胞增殖活性,浓度为100ng/ml时刺激活性更强.结论表达并复性后的重组人SCF-TPO融合蛋白具有刺激MO7e细胞生长活性.
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