Inflammasomes are multiprotein intracellular complexes which are responsible for the activation of inflammatory responses. Among various subtypes of inflammasomes, NLRP3 has been a subject of intensive investigation. ...Inflammasomes are multiprotein intracellular complexes which are responsible for the activation of inflammatory responses. Among various subtypes of inflammasomes, NLRP3 has been a subject of intensive investigation. NLRP3 is considered to be a sensor of microbial and other danger signals and plays a crucial role in mucosal immune responses, promoting the maturation of proinflammatory cytokines interleukin 1β(IL-1β) and IL-18. NLRP3 inflammasome has been associated with a variety of inflammatory and autoimmune conditions, including inflammatory bowel diseases(IBD). The role of NLRP3 in IBD is not yet fully elucidated as it seems to demonstrate both pathogenic and protective effects. Studies have shown a relationship between genetic variants and mutations in NLRP3 gene with IBD pathogenesis. A complex interaction between the NLRP3 inflammasome and the mucosal immune response has been reported. Activation of the inflammasome is a key function mediated by the innate immune response and in parallel the signaling through IL-1β and IL-18 is implicated in adaptive immunity. Further research is needed to delineate the precise mechanisms of NLRP3 function in regulating immune responses. Targeting NLRP3 inflammasome and its downstream signaling will provide new insights into the development of future therapeutic strategies.展开更多
以亚洲百合品种Pollyanna和东方百合品种Sorbonne为试材,在比较异硫氰酸胍法、SDS/酚法与CTAB-L i Cl法提取总RNA效果的基础上,针对百合组织中富含多糖的特点,在Sorbonne提取RNA中加入特殊除多糖步骤,改进了CTAB- L i Cl法.结果表明,改...以亚洲百合品种Pollyanna和东方百合品种Sorbonne为试材,在比较异硫氰酸胍法、SDS/酚法与CTAB-L i Cl法提取总RNA效果的基础上,针对百合组织中富含多糖的特点,在Sorbonne提取RNA中加入特殊除多糖步骤,改进了CTAB- L i Cl法.结果表明,改进CTAB- L i Cl法能有效去除多糖,提取到的RNA2 8S r RNA亮度约为18S r RNA的2倍,A2 6 0 / 2 80介于1.8～2 .0之间,A2 6 0 / 2 30为2 .0 ,Pollyanna RNA产率为36 .3μL·g- 1 ,Sor-bonne RNA产率为10 .2 μL·g- 1 ,经RT- PCR获得了特异性条带,说明用改进CTAB- L i Cl法从百合花瓣中提取到的RNA质量好、产率高、完整性强,完全适合于进一步的分子生物学研究.展开更多
选择合适的内参基因是提高实时荧光定量PCR分析(q RT-PCR)准确性的重要条件。18S r RNA基因表达范围广、表达量恒定,常作为内参基因应用于实时荧光定量PCR中。为了获得丝瓜18S r RNA基因,并设计合适的荧光定量PCR内参引物,解决丝瓜实时...选择合适的内参基因是提高实时荧光定量PCR分析(q RT-PCR)准确性的重要条件。18S r RNA基因表达范围广、表达量恒定,常作为内参基因应用于实时荧光定量PCR中。为了获得丝瓜18S r RNA基因,并设计合适的荧光定量PCR内参引物,解决丝瓜实时荧光定量PCR检测中无内参基因的现状,通过PCR和序列测定,首次克隆到了丝瓜的18S r RNA基因序列,其长度为1 862 bp,Gen Bank登录号为KM656452。在此基础上设计1对荧光定量PCR引物,该引物特异性强,扩增效率高,在丝瓜各生长发育阶段及各种非生物胁迫条件下均能稳定表达,适合在丝瓜基因表达研究中作为内参基因。该研究结果可为开展丝瓜重要功能基因的表达模式和调控机制的研究奠定基础。展开更多
文摘Inflammasomes are multiprotein intracellular complexes which are responsible for the activation of inflammatory responses. Among various subtypes of inflammasomes, NLRP3 has been a subject of intensive investigation. NLRP3 is considered to be a sensor of microbial and other danger signals and plays a crucial role in mucosal immune responses, promoting the maturation of proinflammatory cytokines interleukin 1β(IL-1β) and IL-18. NLRP3 inflammasome has been associated with a variety of inflammatory and autoimmune conditions, including inflammatory bowel diseases(IBD). The role of NLRP3 in IBD is not yet fully elucidated as it seems to demonstrate both pathogenic and protective effects. Studies have shown a relationship between genetic variants and mutations in NLRP3 gene with IBD pathogenesis. A complex interaction between the NLRP3 inflammasome and the mucosal immune response has been reported. Activation of the inflammasome is a key function mediated by the innate immune response and in parallel the signaling through IL-1β and IL-18 is implicated in adaptive immunity. Further research is needed to delineate the precise mechanisms of NLRP3 function in regulating immune responses. Targeting NLRP3 inflammasome and its downstream signaling will provide new insights into the development of future therapeutic strategies.
文摘以亚洲百合品种Pollyanna和东方百合品种Sorbonne为试材,在比较异硫氰酸胍法、SDS/酚法与CTAB-L i Cl法提取总RNA效果的基础上,针对百合组织中富含多糖的特点,在Sorbonne提取RNA中加入特殊除多糖步骤,改进了CTAB- L i Cl法.结果表明,改进CTAB- L i Cl法能有效去除多糖,提取到的RNA2 8S r RNA亮度约为18S r RNA的2倍,A2 6 0 / 2 80介于1.8～2 .0之间,A2 6 0 / 2 30为2 .0 ,Pollyanna RNA产率为36 .3μL·g- 1 ,Sor-bonne RNA产率为10 .2 μL·g- 1 ,经RT- PCR获得了特异性条带,说明用改进CTAB- L i Cl法从百合花瓣中提取到的RNA质量好、产率高、完整性强,完全适合于进一步的分子生物学研究.
文摘选择合适的内参基因是提高实时荧光定量PCR分析(q RT-PCR)准确性的重要条件。18S r RNA基因表达范围广、表达量恒定,常作为内参基因应用于实时荧光定量PCR中。为了获得丝瓜18S r RNA基因,并设计合适的荧光定量PCR内参引物,解决丝瓜实时荧光定量PCR检测中无内参基因的现状,通过PCR和序列测定,首次克隆到了丝瓜的18S r RNA基因序列,其长度为1 862 bp,Gen Bank登录号为KM656452。在此基础上设计1对荧光定量PCR引物,该引物特异性强,扩增效率高,在丝瓜各生长发育阶段及各种非生物胁迫条件下均能稳定表达,适合在丝瓜基因表达研究中作为内参基因。该研究结果可为开展丝瓜重要功能基因的表达模式和调控机制的研究奠定基础。
