目的对细粒棘球蚴抗原B(EgAgB)的3个亚单位(EgAgB1、EgAgB2和EgAgB4)的反应性表位的结构进行预测分析并进行基因重组,鉴定构建的多表位重组抗原的反应性。方法用Bioedit和Discovery Studio Visualizer分析软件和I-TASSER在线服务器对EgA...目的对细粒棘球蚴抗原B(EgAgB)的3个亚单位(EgAgB1、EgAgB2和EgAgB4)的反应性表位的结构进行预测分析并进行基因重组,鉴定构建的多表位重组抗原的反应性。方法用Bioedit和Discovery Studio Visualizer分析软件和I-TASSER在线服务器对EgAgB1、EgAgB2和EgAgB4等3个亚单位抗原序列及其不同组合方式的组合序列进行分析和结构预测。选择结构预测评分较高的表位或亚单位组合方式进行基因重组。设计特异性重叠引物,用重叠延伸PCR技术扩增目标序列。将目标序列克隆至pET32a(+)载体中构建表达质粒表达重组蛋白,表达产物经纯化后即为多表位重组抗原。采用蛋白质印迹(Western blotting)分析鉴定多表位重组抗原的反应性。结果结构预测显示,EgAgB1、EgAgB2和EgAgB4等3个亚单位的主要表位均呈"Z"字型结构,且主要表位区域亦均位于序列的中部;对拟进行组合的3个EgAgB亚单位和4个主要表位(KK36、RK30、B4-2和B4-3)的57种不同的组合方式进行了结构预测,选择6种组合方式进行重组表达。对6个多表位重组抗原(MEA-8、MEA-20、MEA-26、MEA-36、MEA-49和MEA-52)进行的Western blotting分析显示,多表位重组抗原与细粒棘球蚴病患者血清的反应条带明显强于AgB亚单位抗原。结论构建的6个多表位重组抗原与细粒棘球蚴病患者血清的反应性明显强于EgAgB亚单位抗原。展开更多
Transient receptor potential(TRP)channels are widely found throughout the animal kingdom.By serving as cellular sensors for a wide spectrum of physical and chemical stimuli,they play crucial physiological roles rangin...Transient receptor potential(TRP)channels are widely found throughout the animal kingdom.By serving as cellular sensors for a wide spectrum of physical and chemical stimuli,they play crucial physiological roles ranging from sensory transduction to cell cycle modulation.TRP channels are tetrameric protein complexes.While most TRP subunits can form functional homomeric channels,heteromerization of TRP channel subunits of either the same subfamily or different subfamilies has been widely observed.Heteromeric TRP channels exhibit many novel properties compared to their homomeric counterparts,indicating that co-assembly of TRP channel subunits has an important contribution to the diversity of TRP channel functions.展开更多
文摘目的对细粒棘球蚴抗原B(EgAgB)的3个亚单位(EgAgB1、EgAgB2和EgAgB4)的反应性表位的结构进行预测分析并进行基因重组,鉴定构建的多表位重组抗原的反应性。方法用Bioedit和Discovery Studio Visualizer分析软件和I-TASSER在线服务器对EgAgB1、EgAgB2和EgAgB4等3个亚单位抗原序列及其不同组合方式的组合序列进行分析和结构预测。选择结构预测评分较高的表位或亚单位组合方式进行基因重组。设计特异性重叠引物,用重叠延伸PCR技术扩增目标序列。将目标序列克隆至pET32a(+)载体中构建表达质粒表达重组蛋白,表达产物经纯化后即为多表位重组抗原。采用蛋白质印迹(Western blotting)分析鉴定多表位重组抗原的反应性。结果结构预测显示,EgAgB1、EgAgB2和EgAgB4等3个亚单位的主要表位均呈"Z"字型结构,且主要表位区域亦均位于序列的中部;对拟进行组合的3个EgAgB亚单位和4个主要表位(KK36、RK30、B4-2和B4-3)的57种不同的组合方式进行了结构预测,选择6种组合方式进行重组表达。对6个多表位重组抗原(MEA-8、MEA-20、MEA-26、MEA-36、MEA-49和MEA-52)进行的Western blotting分析显示,多表位重组抗原与细粒棘球蚴病患者血清的反应条带明显强于AgB亚单位抗原。结论构建的6个多表位重组抗原与细粒棘球蚴病患者血清的反应性明显强于EgAgB亚单位抗原。
文摘Transient receptor potential(TRP)channels are widely found throughout the animal kingdom.By serving as cellular sensors for a wide spectrum of physical and chemical stimuli,they play crucial physiological roles ranging from sensory transduction to cell cycle modulation.TRP channels are tetrameric protein complexes.While most TRP subunits can form functional homomeric channels,heteromerization of TRP channel subunits of either the same subfamily or different subfamilies has been widely observed.Heteromeric TRP channels exhibit many novel properties compared to their homomeric counterparts,indicating that co-assembly of TRP channel subunits has an important contribution to the diversity of TRP channel functions.
