无细胞蛋白质表达系统(cell-free protein expression systems,CFPS)是一种快速发展的蛋白质表达系统,由细胞裂解物、能源物质、底物和外源模板在细胞外转录、翻译和折叠来合成目的蛋白质。与传统体内表达体系相比,无细胞表达系统没有...无细胞蛋白质表达系统(cell-free protein expression systems,CFPS)是一种快速发展的蛋白质表达系统,由细胞裂解物、能源物质、底物和外源模板在细胞外转录、翻译和折叠来合成目的蛋白质。与传统体内表达体系相比,无细胞表达系统没有细胞生长过程,没有细胞壁阻碍,可操作性强,是探索蛋白质遗传密码、制备特殊蛋白、开展高通量药物筛选和蛋白质组学研究的重要工具。本文介绍了无细胞蛋白质表达系统的不同类型,分析了影响体系活力的原因,并阐述了优化体系表达量的方法和策略。文章对CFPS的优化中运用新兴材料与基因模板相结合提高蛋白质表达量的技术进行了详细论述,并总结了无细胞蛋白质表达系统的实际应用,显示了其在合成蛋白质领域的重要潜力。展开更多
Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage ...Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage line U937 (as control) and U937 with MCP-1 was extracted, made reverse transcript to cDNA and tested with gene expression chip HO2 human. Results: Some chemotactic-related gene expressions were changed in all analyzed genes. Regulated upon activation, normal T cell expressed and secreted (RANTES) was up-regulated over 2-fold and 7 chemotactic-related genes (CCR2, CCR5, CCL16, GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2) were down-regulated over 2-fold in MCP-1 treated U937 cells at mRNA level. Conclusion: MCP-1 can influence some chemokines and receptors expression in macrophage in vitro, in which MCP-1 mainly down-regulates the chemotactic genes expression of those influencing neutrophilic granulocyte (GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2). Another novel finding is that it can also down-regulate the mRNA level of CCR5, which plays a critical role in many disorders and illnesses.展开更多
文摘无细胞蛋白质表达系统(cell-free protein expression systems,CFPS)是一种快速发展的蛋白质表达系统,由细胞裂解物、能源物质、底物和外源模板在细胞外转录、翻译和折叠来合成目的蛋白质。与传统体内表达体系相比,无细胞表达系统没有细胞生长过程,没有细胞壁阻碍,可操作性强,是探索蛋白质遗传密码、制备特殊蛋白、开展高通量药物筛选和蛋白质组学研究的重要工具。本文介绍了无细胞蛋白质表达系统的不同类型,分析了影响体系活力的原因,并阐述了优化体系表达量的方法和策略。文章对CFPS的优化中运用新兴材料与基因模板相结合提高蛋白质表达量的技术进行了详细论述,并总结了无细胞蛋白质表达系统的实际应用,显示了其在合成蛋白质领域的重要潜力。
文摘Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage line U937 (as control) and U937 with MCP-1 was extracted, made reverse transcript to cDNA and tested with gene expression chip HO2 human. Results: Some chemotactic-related gene expressions were changed in all analyzed genes. Regulated upon activation, normal T cell expressed and secreted (RANTES) was up-regulated over 2-fold and 7 chemotactic-related genes (CCR2, CCR5, CCL16, GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2) were down-regulated over 2-fold in MCP-1 treated U937 cells at mRNA level. Conclusion: MCP-1 can influence some chemokines and receptors expression in macrophage in vitro, in which MCP-1 mainly down-regulates the chemotactic genes expression of those influencing neutrophilic granulocyte (GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2). Another novel finding is that it can also down-regulate the mRNA level of CCR5, which plays a critical role in many disorders and illnesses.
