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目的:构建hSCGF-α穿膜肽(TAT-SCGF),研究其对人脐带间充质干细胞的增殖作用。方法:采用基因工程技术将hSCGF-α基因重组插入pET-28a(+)质粒DNA,构建TAT-SCGF-28a重组DNA,并在大肠杆菌BL21(DE3)中成功高效表达,包涵体纯化后,通过实验比较hSCGF-α及hSCGF-α穿膜肽(TAT-SCGF)对人脐带间充质干细胞的增殖刺激作用。结果与结论:成功构建TAT-SCGF-28a重组DNA;纯化的蛋白纯度达90%以上;TAT-SCGF较不具有穿膜功能的SCGF对人脐带间充质干细胞有更强的增殖效果。
OBJECTIVE: To construct hSCGF-α transmembrane peptide (TAT-SCGF) and study its proliferation on human umbilical cord mesenchymal stem cells. Methods: Recombinant plasmid pET-28a (+) was inserted into recombinant plasmid pET-28a (+) to construct recombinant plasmid TAT-SCGF-28a. The recombinant plasmid was successfully expressed in E.coli BL21 (DE3) The effects of hSCGF-α and hSCGF-α transmembrane peptide (TAT-SCGF) on the proliferation of human umbilical cord mesenchymal stem cells were compared. RESULTS AND CONCLUSION: TAT-SCGF-28a recombinant DNA was successfully constructed. The purity of the purified protein was above 90%. Compared with the non-transmembrane-functionalized SCGF, TAT-SCGF could enhance the proliferation of human umbilical cord mesenchymal stem cells.