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重组GPI-B7胞外段在中华仓鼠卵巢癌细胞膜上的表达
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作者 程绍辉 于继云 +1 位作者 陈兴 许红莲 《免疫学杂志》 CAS CSCD 北大核心 2003年第S1期1-4,共4页
目的 通过从衰变加速因子(DAF)来源的GPI修饰性信号序列克隆到PCI-dhfr真核表达载体,利用GPI信号序列对B7-1基因胞外段序列进行修饰,实现GPI-B7-1锚定蛋白在中华仓鼠卵巢癌细胞(Chinese hamster ovary cell,CHO)膜表面的表达。方法 应用... 目的 通过从衰变加速因子(DAF)来源的GPI修饰性信号序列克隆到PCI-dhfr真核表达载体,利用GPI信号序列对B7-1基因胞外段序列进行修饰,实现GPI-B7-1锚定蛋白在中华仓鼠卵巢癌细胞(Chinese hamster ovary cell,CHO)膜表面的表达。方法 应用RT-PCR方法从B淋巴细胞瘤细胞系(Raji细胞系)总RNA中钓取B7-1(CD80)全长基因,扩增B7-1胞外段分子并克隆到已含有GPI修饰性信号序列的真核表达载体pCI-dhfr上,应用脂质体方法转染到CHO-dhfr-细胞中,用氨甲喋呤(Methotrexate,MTX)进行筛选。重组蛋白的表达用细胞免疫荧光进行鉴定。结果 成功地克隆了B7-1全长基因,并扩增了胞外段基因构建到含有GPI修饰性信号序列的真核表达载体pCI-dhfr上,实现了GPI-B7-1锚定蛋白在CHO细胞膜的表达。结论GPI信号序列能够通过其脂质性尾部将GPI-B7-1锚定蛋白整合到细胞膜表面,本研究结果为将来应用GPI-B7-1分子于肿瘤细胞膜表面的修饰做了技术准备,可作为肿瘤免疫治疗的手段。 展开更多
关键词 RT-PCR B7-1 gpi修饰蛋白 膜表达
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A novel vWF A3-GPI modified EPCs to enhance its adhesion ability to collegen 被引量:1
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作者 Tan Hu Song Yaoming Jin Jun Zhao Xiaohui Huang Lan 《Journal of Medical Colleges of PLA(China)》 CAS 2011年第4期191-203,共13页
Objective:To gain purified recombination protein rGPI-vWF A3 which can guide endothelial progenitor cells (EPCs) gathering to the internal surface of the injured vessels theoretically and analyze its bioactivity in vi... Objective:To gain purified recombination protein rGPI-vWF A3 which can guide endothelial progenitor cells (EPCs) gathering to the internal surface of the injured vessels theoretically and analyze its bioactivity in vitro. Methods: The plasmid vector of vWF A3 fusion protein which contained GPI structure was constructed and recombination protein rGPI-vWF A3 was expressed and its bioactivity was analyzed by ELISA and FCM. Results: Fusion protein vWF A3-GPI was expressed and purified by immunoaffinity chromatography and it could stably anchored on the surface of EPCs and showed its biological activities of collagen conjunction. Conclusion: These findings represent a novel strategy in EPC transplantation treatment for vessel injury. 展开更多
关键词 vWF A3-gpi Protein endothelial progenitor cells COLLAGEN
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