采用毕赤酵母系统对粪肠球菌核糖体蛋白S11(ribosomal protein S11,RPS11)进行异源表达。通过优化RPS11基因序列以适应酵母的密码子偏好,构建重组表达载体pPIC9K-RPS11;采用电击法将其转化至毕赤酵母GS115菌株,并通过G418抗性筛选获得...采用毕赤酵母系统对粪肠球菌核糖体蛋白S11(ribosomal protein S11,RPS11)进行异源表达。通过优化RPS11基因序列以适应酵母的密码子偏好,构建重组表达载体pPIC9K-RPS11;采用电击法将其转化至毕赤酵母GS115菌株,并通过G418抗性筛选获得高拷贝重组菌株;采用甲醇诱导蛋白表达,并通过SDS-PAGE验证了表达效果;将重组蛋白应用于小鼠黑色素瘤治疗模型,以分析其治疗效果。结果显示:重组表达载体pPIC9K-RPS11成功在毕赤酵母中表达了相对分子质量约为14 kDa的目的蛋白;发酵条件为摇瓶诱导温度30℃、诱导时间72 h、甲醇浓度为1%;小鼠腹腔巨噬细胞驯化免疫模型分析表明,重组RPS11具有诱导驯化免疫的生物学活性;小鼠黑色素瘤模型的治疗试验显示,重组RPS11能够显著抑制肿瘤生长。结果表明,毕赤酵母分泌表达的重组RPS11不仅具有诱导驯化免疫的生物学活性,而且对小鼠黑色素瘤具有抑制肿瘤细胞生长的作用,这为异源表达重组RPS11及其进一步应用提供了理论依据。展开更多
The high Li-ion conductivity of the Li7P3S11 sulfide-based solid electrolyte makes it a promising candidate for all-solid-state lithium batteries. The Li-ion transport over electrode-electrolyte and electrolyteelectro...The high Li-ion conductivity of the Li7P3S11 sulfide-based solid electrolyte makes it a promising candidate for all-solid-state lithium batteries. The Li-ion transport over electrode-electrolyte and electrolyteelectrolyte interfaces, vital for the performance of solid-state batteries, is investigated by impedance spectroscopy and solid-state NMR experiments. An all-solid-state Li-ion battery is assembled with the Li7P3S11 electrolyte, nano-Li2S cathode and Li-In foil anode, showing a relatively large initial discharge capacity of 1139.5 m Ah/g at a current density of 0.064 m A/cm^ 2 retaining 850.0 m Ah/g after 30 cycles. Electrochemical impedance spectroscopy suggests that the decrease in capacity over cycling is due to the increased interfacial resistance between the electrode and the electrolyte. 1D exchange ^7Li NMR quantifies the interfacial Li-ion transport between the uncycled electrode and the electrolyte, resulting in a diffusion coefficient of 1.70(3) ×10^-14cm^2/s at 333 K and an energy barrier of 0.132 e V for the Li-ion transport between Li2S cathode and Li7P3S11 electrolyte. This indicates that the barrier for Li-ion transport over the electrode-electrolyte interface is small. However, the small diffusion coefficient for Li-ion diffusion between the Li2S and the Li7P3S11 suggests that these contact interfaces between electrode and electrolyte are relatively scarce, challenging the performance of these solid-state batteries.展开更多
文摘采用毕赤酵母系统对粪肠球菌核糖体蛋白S11(ribosomal protein S11,RPS11)进行异源表达。通过优化RPS11基因序列以适应酵母的密码子偏好,构建重组表达载体pPIC9K-RPS11;采用电击法将其转化至毕赤酵母GS115菌株,并通过G418抗性筛选获得高拷贝重组菌株;采用甲醇诱导蛋白表达,并通过SDS-PAGE验证了表达效果;将重组蛋白应用于小鼠黑色素瘤治疗模型,以分析其治疗效果。结果显示:重组表达载体pPIC9K-RPS11成功在毕赤酵母中表达了相对分子质量约为14 kDa的目的蛋白;发酵条件为摇瓶诱导温度30℃、诱导时间72 h、甲醇浓度为1%;小鼠腹腔巨噬细胞驯化免疫模型分析表明,重组RPS11具有诱导驯化免疫的生物学活性;小鼠黑色素瘤模型的治疗试验显示,重组RPS11能够显著抑制肿瘤生长。结果表明,毕赤酵母分泌表达的重组RPS11不仅具有诱导驯化免疫的生物学活性,而且对小鼠黑色素瘤具有抑制肿瘤细胞生长的作用,这为异源表达重组RPS11及其进一步应用提供了理论依据。
基金funding from the European Research Council under the European Union’s Seventh Framework Programme (FP/2007-2013)/ERC Grant Agreement no.[307161] of M.W.
文摘The high Li-ion conductivity of the Li7P3S11 sulfide-based solid electrolyte makes it a promising candidate for all-solid-state lithium batteries. The Li-ion transport over electrode-electrolyte and electrolyteelectrolyte interfaces, vital for the performance of solid-state batteries, is investigated by impedance spectroscopy and solid-state NMR experiments. An all-solid-state Li-ion battery is assembled with the Li7P3S11 electrolyte, nano-Li2S cathode and Li-In foil anode, showing a relatively large initial discharge capacity of 1139.5 m Ah/g at a current density of 0.064 m A/cm^ 2 retaining 850.0 m Ah/g after 30 cycles. Electrochemical impedance spectroscopy suggests that the decrease in capacity over cycling is due to the increased interfacial resistance between the electrode and the electrolyte. 1D exchange ^7Li NMR quantifies the interfacial Li-ion transport between the uncycled electrode and the electrolyte, resulting in a diffusion coefficient of 1.70(3) ×10^-14cm^2/s at 333 K and an energy barrier of 0.132 e V for the Li-ion transport between Li2S cathode and Li7P3S11 electrolyte. This indicates that the barrier for Li-ion transport over the electrode-electrolyte interface is small. However, the small diffusion coefficient for Li-ion diffusion between the Li2S and the Li7P3S11 suggests that these contact interfaces between electrode and electrolyte are relatively scarce, challenging the performance of these solid-state batteries.
