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目的 可溶性人干细胞因子 (SCF)膜外活性区与增强型绿色荧光蛋白 (EGFP)融合基因表达载体的构建及其表达。方法 应用基因工程技术构建重组载体 ,电穿孔转染CHO K1细胞株 ,荧光分光光度计检测上清液中融合蛋白的表达 ,流式细胞仪检测其活性。结果 融合基因在瞬时转染的真核细胞中获得表达 ,并分泌至上清 ,且SCF EGFP融合蛋白中分子标签EGFP未影响可溶性人SCF膜外活性区的结构及其生物学活性。结论 可溶性人干细胞因子与EGFP融合基因载体构建成功 ,表达的融合蛋白具有生物活性 ,为进一步研究SCF对造血干细胞的生物学作用奠定了基础。
Objective To construct and express the fusion gene expression vector of soluble extracellular domain of soluble human stem cell factor (SCF) and enhanced green fluorescent protein (EGFP). Methods Recombinant vector was constructed by gene engineering technology. The CHO K1 cell line was transfected by electroporation. The fusion protein was detected by fluorescence spectrophotometer and the activity of fusion protein was detected by flow cytometry. Results The fusion gene was expressed in transiently transfected eukaryotic cells and secreted into the supernatant. The molecular tagged EGFP in SCF EGFP fusion protein did not affect the structure and biological activity of soluble human SCF. Conclusions The soluble human stem cell factor and EGFP fusion gene vector was successfully constructed and the expressed fusion protein was bioactive, which lays the foundation for further study on the biological effect of SCF on hematopoietic stem cells.