小鼠颗粒酶F重组蛋白表达与功能鉴定
he expression and function of mouse recombinant granzyme F
目的：细胞毒性T淋巴细胞和自然杀伤细胞通过释放毒性颗粒来引起靶细胞的死亡，其中小鼠NK细胞具有特有的颗粒酶F，本文试图表达具有活性的颗粒酶F，并研究其酶活特点。方法：构建不同的表达载体，通过酿酒酵母胞内表达、毕赤酵母分泌表达、昆虫SF21细胞表达以及大肠杆菌表达尝试表达活性颗粒酶F。结果：酿酒酵母、毕赤酵母、昆虫细胞等方式都没有表达出活性的颗粒酶F，最终通过大肠杆菌包涵体变复性的方法，成功的表达纯化了颗粒酶F的蛋白方法，并进行了活化处理和功能鉴定。结论：通过包涵体变复性成功表达出具有活性的颗粒酶F，为下一步的研究奠定了基础。
ytotoxic T lymphocytes(CTL) and natural killer(NK) cells are important effector in virus-infected and tumor cells elimination. Granule exocytosis is the main pathway for the killing of target cells. Granzyme F(GzmF) belongs to a unique group of granzymes in mice. The murine GzmF is highly expressed in NK cells and lymphokine-activated killer (LAK) cells. However, how GzmF works in granule-mediated cytolysis is still unknown. We successfully express and purify GzmF from E.coli inclusion body, identify the specific enzyme activity of granzyme F towards synthetic substrate and discover the cytotoxic activity of GzmF in killing tumor cells.
黄锐、石磊
生物工程学分子生物学生物化学
NK细胞颗粒酶F蛋白表达包涵体
NK cellgranzyme Fprotein expressioninclusion body
黄锐,石磊.小鼠颗粒酶F重组蛋白表达与功能鉴定[EB/OL].(2016-05-17)[2025-08-11].http://www.paper.edu.cn/releasepaper/content/201605-470.点此复制
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