The exogenous gene was integrated into Dunaliella salina successfully by using LiAc/PEG mediating method for the first time. According to the results of histochemical staining, transgenic D. salina was blue, showing t...The exogenous gene was integrated into Dunaliella salina successfully by using LiAc/PEG mediating method for the first time. According to the results of histochemical staining, transgenic D. salina was blue, showing that the exogenous GUS gene was successfully expressed in the cells of D. salina. Meanwhile, the effects of growth state of D. salina, plasmid concentration and temperature on its transformation efficiency were studied, and the transformation conditions were optimized. The results show that the optimum conditions for the genetic transformation of D. salina are shown as follows： D. salina was in the early logarithmic phase; plasmid DNA concentration was 600 μg/ml; temperature was 29 ℃, and transformation efficiency was up to 74.8‰ under the best conditions. According to the results of PCR amplification and PCR-Southern hybridization, the target gene had been integrated into genome and was hereditary.展开更多
目的合成聚乙二醇单甲醚-聚乳酸(mPEG-PDLLA)嵌段共聚物,制备去氧鬼臼毒素mPEG-PDLLA嵌段共聚物(DPT-PM),提高去氧鬼臼毒素在水中的溶解度。方法以开环聚合反应合成mPEG-PDLLA,并通过IR、1H-HMR确证其结构,芘荧光探针法测定其临界胶束浓...目的合成聚乙二醇单甲醚-聚乳酸(mPEG-PDLLA)嵌段共聚物,制备去氧鬼臼毒素mPEG-PDLLA嵌段共聚物(DPT-PM),提高去氧鬼臼毒素在水中的溶解度。方法以开环聚合反应合成mPEG-PDLLA,并通过IR、1H-HMR确证其结构,芘荧光探针法测定其临界胶束浓度(CMC);采用溶剂蒸发法制备DPT-PM溶液,并将其冷冻干燥;分别采用动态光散射法(DLS)、透射电子显微镜(TEM)、差示扫描量热法(DSC)、高效液相色谱法(HPLC)等手段对胶束形态、粒径与分布、载药量、包封率等进行表征,并采用透析法考察DPT-PM体外释放,并对释放机制进行探讨。结果制备了去氧鬼臼毒素共聚物胶束,透射电镜下观察为近球形,平均粒径为22.0±8.9nm,载药量为20.67%,包封率为99.63%。结论 m PEG-PDLLA聚合物胶束可作为疏水性药物去氧鬼臼毒素的载体,具有较高的载药性能,能一定程度提高去氧鬼臼毒素在水中的溶解度。展开更多
基金Supported by National Natural Science Foundation of China(31472260)~~
文摘The exogenous gene was integrated into Dunaliella salina successfully by using LiAc/PEG mediating method for the first time. According to the results of histochemical staining, transgenic D. salina was blue, showing that the exogenous GUS gene was successfully expressed in the cells of D. salina. Meanwhile, the effects of growth state of D. salina, plasmid concentration and temperature on its transformation efficiency were studied, and the transformation conditions were optimized. The results show that the optimum conditions for the genetic transformation of D. salina are shown as follows： D. salina was in the early logarithmic phase; plasmid DNA concentration was 600 μg/ml; temperature was 29 ℃, and transformation efficiency was up to 74.8‰ under the best conditions. According to the results of PCR amplification and PCR-Southern hybridization, the target gene had been integrated into genome and was hereditary.
文摘目的合成聚乙二醇单甲醚-聚乳酸(mPEG-PDLLA)嵌段共聚物,制备去氧鬼臼毒素mPEG-PDLLA嵌段共聚物(DPT-PM),提高去氧鬼臼毒素在水中的溶解度。方法以开环聚合反应合成mPEG-PDLLA,并通过IR、1H-HMR确证其结构,芘荧光探针法测定其临界胶束浓度(CMC);采用溶剂蒸发法制备DPT-PM溶液,并将其冷冻干燥;分别采用动态光散射法(DLS)、透射电子显微镜(TEM)、差示扫描量热法(DSC)、高效液相色谱法(HPLC)等手段对胶束形态、粒径与分布、载药量、包封率等进行表征,并采用透析法考察DPT-PM体外释放,并对释放机制进行探讨。结果制备了去氧鬼臼毒素共聚物胶束,透射电镜下观察为近球形,平均粒径为22.0±8.9nm,载药量为20.67%,包封率为99.63%。结论 m PEG-PDLLA聚合物胶束可作为疏水性药物去氧鬼臼毒素的载体,具有较高的载药性能,能一定程度提高去氧鬼臼毒素在水中的溶解度。
