sflk1-IFN-γ融合蛋白基因重组质粒的构建、表达及生物学活性鉴定

发布时间：2018-05-22 16:30

本文选题：血管内皮生长因子受体2 + 融合蛋白　；参考：《浙江大学》2006年硕士论文

【摘要】：恶性肿瘤的生长、浸润和转移必须依靠肿瘤新生血管提供足够的营养,因此抑制或破坏肿瘤血管生成已成为近年来肿瘤治疗的新方法。血管内皮生长因子(VEGF)与表达在血管内皮细胞上的相应受体VEGFR2结合所引起的信号转导是血管生成中的限速步骤,在整个血管生成中发挥着关键作用。可溶性血管内皮生长因子受体2(sVEGFR2,在小鼠中被称为sflkl)可竞争性地结合VEGF,从而阻断VEGF与VEGFR2结合所引起的信号转导,抑制肿瘤细胞诱导的血管生成和肿瘤的生长。IFN-γ是细胞介导的免疫应答中非常重要的效应因子,能诱导CTL应答及Th1细胞的分化,并且尚能直接抑制多种肿瘤的生长,同时它自身也有抑制肿瘤血管生成的作用。因此我们构建了pcDNA3.1(+)/sflk1-IFN-γ重组质粒,将其在CHO细胞中高效稳定表达,并对其表达产物sflk1-IFN-γ融合蛋白的生物学活性进行了研究鉴定。目的构建小鼠可溶性的血管内皮生长因子受体2(sflkl)与小鼠IFN-γ(mIFN-γ)融合基因的真核表达质粒pcDNA3.1(+)/silk1—IFN-γ;将该重组质粒在CHO细胞中表达,筛选出高效稳定表达sflk1-IFN-γ融合蛋白的CHO细胞株;并对该融合蛋白的生物学活性进行鉴定。
[Abstract]:The growth, invasion and metastasis of malignant tumors must rely on tumor neovascularization to provide adequate nutrition, so inhibition or destruction of tumor angiogenesis has become a new method of tumor treatment in recent years. The signal transduction caused by the binding of vascular endothelial growth factor (VEGF) to the corresponding receptor VEGFR2 expressed on vascular endothelial cells is a rate-limiting step in angiogenesis and plays a key role in the whole angiogenesis. Soluble vascular endothelial growth factor receptor 2sVEGFR2, known as sflkl2 in mice, can competitively bind to VEGF, thus blocking the signal transduction caused by the binding of VEGF to VEGFR2. Inhibiting angiogenesis induced by tumor cells and tumor growth. IFN- 纬 is a very important effector factor in cellular mediated immune response, which can induce CTL response and differentiation of Th1 cells, and can directly inhibit the growth of many kinds of tumors. At the same time, it also has the effect of inhibiting tumor angiogenesis. The recombinant plasmid pcDNA3.1 (/ sflk1-IFN- 纬) was constructed and expressed stably in CHO cells, and the biological activity of the fusion protein sflk1-IFN- 纬 was studied. Purpose The eukaryotic expression plasmid pcDNA3.1 (r-silk1-IFN- 纬) of the fusion gene of soluble vascular endothelial growth factor receptor (sflkl2) and mouse IFN- 纬 was constructed, and the recombinant plasmid was expressed in CHO cells to screen the CHO cell line with high and stable expression of sflk1-IFN- 纬 fusion protein. The biological activity of the fusion protein was identified.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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