Objective To construct the human interleukin 18 DNA plasmids vaccine and to express the eukaryotic plasmids vaccine in mammalian cell lines Cos 7 and D5.Methods Gene recombinant technique was used to construct hIL 18 ...Objective To construct the human interleukin 18 DNA plasmids vaccine and to express the eukaryotic plasmids vaccine in mammalian cell lines Cos 7 and D5.Methods Gene recombinant technique was used to construct hIL 18 eukaryotic expression vectors.Calcium phosphate method was performed to transect recombinant hIL 18 eukaryotic expression vectors into Cos 7 and D5 cells. In situ hybridization and Western Blot were implemented to verify the transient expression of recombinant hIL 18 in Cos 7 and D5.Results The eukaryotic expression plasmid pVAX1 IL 18 was constructed successfully.hIL 18 was transiently expressed in Cos 7 and D5.Conclusion The eukaryotic expression plasmid pVAX1 IL 18 was constructed. In situ hybridization and Western Blot results proved the successful transient expression of pVAX1 IL 18 in Cos 7 and D5.Therefore,the work has settled the foundation for further biological research on hIL 18,including immunogene therapy through hIL 18.展开更多
以牡丹籽壳为原料,利用大孔树脂和C18反相键合硅胶柱(C18柱)联合分离短叶松素,回收率为(98.33±0.64)%。用70%(体积分数)乙醇提取牡丹籽壳粗黄酮(Mu Dan Ke Flavonoids,MDKF),得率最高为(10.54±0.13)%;比较了6种大孔树脂(AB-8...以牡丹籽壳为原料,利用大孔树脂和C18反相键合硅胶柱(C18柱)联合分离短叶松素,回收率为(98.33±0.64)%。用70%(体积分数)乙醇提取牡丹籽壳粗黄酮(Mu Dan Ke Flavonoids,MDKF),得率最高为(10.54±0.13)%;比较了6种大孔树脂(AB-8、S-8、DM301、HPD600、HPD100和D101)的吸附率和解析率,发现S-8大孔树脂的吸附率和解析率最佳,分别为83.47%和84.46%;优化S-8大孔树脂分离MDKF的最佳条件为:上样液质量浓度1.6 mg/mL,上样液流速2.0 mL/min,洗脱剂乙醇体积分数60%,洗脱液流速1.5 mL/min,洗脱液体积100 mL;对经过C18柱分离后的组分进行LC-MS分析得到短叶松素,回收率为(98.33±0.64)%;分离前后抗氧化能力比较:C18纯化物>S-8大孔树脂纯化物>MDKF粗提物;分子对接实验表明,短叶松素与超氧化物歧化酶、过氧化氢酶、过氧化物酶、谷胱甘肽过氧化物酶均有结合能力,过氧化氢酶的结合能力最强为-9.1 kcal/mol。实验表明,短叶松素具有较好的抗氧化能力,可作为食品、药品或化妆品等抗氧化剂的添加,应用前景非常广泛,对牡丹籽壳的进一步开发利用提供理论支持。展开更多
文摘Objective To construct the human interleukin 18 DNA plasmids vaccine and to express the eukaryotic plasmids vaccine in mammalian cell lines Cos 7 and D5.Methods Gene recombinant technique was used to construct hIL 18 eukaryotic expression vectors.Calcium phosphate method was performed to transect recombinant hIL 18 eukaryotic expression vectors into Cos 7 and D5 cells. In situ hybridization and Western Blot were implemented to verify the transient expression of recombinant hIL 18 in Cos 7 and D5.Results The eukaryotic expression plasmid pVAX1 IL 18 was constructed successfully.hIL 18 was transiently expressed in Cos 7 and D5.Conclusion The eukaryotic expression plasmid pVAX1 IL 18 was constructed. In situ hybridization and Western Blot results proved the successful transient expression of pVAX1 IL 18 in Cos 7 and D5.Therefore,the work has settled the foundation for further biological research on hIL 18,including immunogene therapy through hIL 18.
文摘以牡丹籽壳为原料,利用大孔树脂和C18反相键合硅胶柱(C18柱)联合分离短叶松素,回收率为(98.33±0.64)%。用70%(体积分数)乙醇提取牡丹籽壳粗黄酮(Mu Dan Ke Flavonoids,MDKF),得率最高为(10.54±0.13)%;比较了6种大孔树脂(AB-8、S-8、DM301、HPD600、HPD100和D101)的吸附率和解析率,发现S-8大孔树脂的吸附率和解析率最佳,分别为83.47%和84.46%;优化S-8大孔树脂分离MDKF的最佳条件为:上样液质量浓度1.6 mg/mL,上样液流速2.0 mL/min,洗脱剂乙醇体积分数60%,洗脱液流速1.5 mL/min,洗脱液体积100 mL;对经过C18柱分离后的组分进行LC-MS分析得到短叶松素,回收率为(98.33±0.64)%;分离前后抗氧化能力比较:C18纯化物>S-8大孔树脂纯化物>MDKF粗提物;分子对接实验表明,短叶松素与超氧化物歧化酶、过氧化氢酶、过氧化物酶、谷胱甘肽过氧化物酶均有结合能力,过氧化氢酶的结合能力最强为-9.1 kcal/mol。实验表明,短叶松素具有较好的抗氧化能力,可作为食品、药品或化妆品等抗氧化剂的添加,应用前景非常广泛,对牡丹籽壳的进一步开发利用提供理论支持。
