The receptor for activated C-kinase 1 （RACK1） is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic rice plants, in ...The receptor for activated C-kinase 1 （RACK1） is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic rice plants, in which the expression of RACK1 gene was inhibited by RNA interference （RNAi）, were studied to elucidate the possible functions of RACK1 in responses to drought stress in rice. Real-time PCR analysis showed that the expression of RACK1 in transgenic rice plants was inhibited by more than 50%. The tolerance to drought stress of the transgenic rice plants was higher as compared with the non-transgenic rice plants. The peroxidation of membrane and the production of malondialdehyde were significantly lower and the superoxide dismutase activity in transgenic rice plants was significantly higher than those in non-trangenic rice plants It is suggested that RACK1 negatively regulated the redox system-related tolerance to drought stress of rice plants.展开更多
目的探讨非诺贝特对人肝瘤细胞株(HepG2)细胞1型纤维酶原激活物抑制剂(PAI-1)表达的影响及机制。方法用不同浓度非诺贝特刺激HepG2细胞,采用半定量逆转录聚合酶链反应(RT-PCR)法检测PAI-1mRNA水平,发色底物法检测PAI-1的活性变化。构建...目的探讨非诺贝特对人肝瘤细胞株(HepG2)细胞1型纤维酶原激活物抑制剂(PAI-1)表达的影响及机制。方法用不同浓度非诺贝特刺激HepG2细胞,采用半定量逆转录聚合酶链反应(RT-PCR)法检测PAI-1mRNA水平,发色底物法检测PAI-1的活性变化。构建4个荧光素酶报告基因质粒,分别由PAI-1启动子序列从-804至+17间不同长度片段驱动,体外转染HepG2细胞,检测荧光素酶的活性。结果非诺贝特能使HepG2细胞PAI-1mRNA表达及蛋白活性显著降低,且呈一定剂量依赖性;还可使PAI-1转录活性显著降低;当转染质粒含有PAI-1启动子序列-636^+17、-449^+17-、276^+17 bp 3个片段时,荧光素酶活性显著增高;共转染过氧化体增殖物激活型受体α(PPARα)表达质粒(PPAR-αpSG5)的细胞在非诺贝特诱导下PAI-1转录活性显著降低。结论非诺贝特可以抑制HepG2细胞PAI-1mRNA表达及其活性,调节PAI-1的基因转录,PPARα参与非诺贝特对PAI-1基因的表达调控。展开更多
鸡核受体辅激活蛋白1基因(NCOA1)mRNA的表达对性腺的发育、成熟均起关键作用,并且与繁殖性状相关。为了建立荧光定量PCR技术检测鸡NCOA1表达量的方法,根据GenBank中NCOA1基因序列设计合成了引物,以β-action基因为内参基因,对荧光定量PC...鸡核受体辅激活蛋白1基因(NCOA1)mRNA的表达对性腺的发育、成熟均起关键作用,并且与繁殖性状相关。为了建立荧光定量PCR技术检测鸡NCOA1表达量的方法,根据GenBank中NCOA1基因序列设计合成了引物,以β-action基因为内参基因,对荧光定量PCR方法进行了评估。结果表明:构建的标准曲线线性关系良好,建立的NCOA1基因荧光定量PCR检测方法灵敏度高、特异性强,准确可靠。建立的SYBR Green I实时荧光定量PCR方法可以测定鸡NCOA1mRNA表达水平的相对含量。展开更多
基金supported by the National Natural Science Foundation of China (Grant No. 30571120)the National High Technology Research and Development Program of China (Grant No.2008AA10Z120)the Research Fund for the Doctoral Program of Higher Education, China
文摘The receptor for activated C-kinase 1 （RACK1） is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic rice plants, in which the expression of RACK1 gene was inhibited by RNA interference （RNAi）, were studied to elucidate the possible functions of RACK1 in responses to drought stress in rice. Real-time PCR analysis showed that the expression of RACK1 in transgenic rice plants was inhibited by more than 50%. The tolerance to drought stress of the transgenic rice plants was higher as compared with the non-transgenic rice plants. The peroxidation of membrane and the production of malondialdehyde were significantly lower and the superoxide dismutase activity in transgenic rice plants was significantly higher than those in non-trangenic rice plants It is suggested that RACK1 negatively regulated the redox system-related tolerance to drought stress of rice plants.
文摘目的探讨非诺贝特对人肝瘤细胞株(HepG2)细胞1型纤维酶原激活物抑制剂(PAI-1)表达的影响及机制。方法用不同浓度非诺贝特刺激HepG2细胞,采用半定量逆转录聚合酶链反应(RT-PCR)法检测PAI-1mRNA水平,发色底物法检测PAI-1的活性变化。构建4个荧光素酶报告基因质粒,分别由PAI-1启动子序列从-804至+17间不同长度片段驱动,体外转染HepG2细胞,检测荧光素酶的活性。结果非诺贝特能使HepG2细胞PAI-1mRNA表达及蛋白活性显著降低,且呈一定剂量依赖性;还可使PAI-1转录活性显著降低;当转染质粒含有PAI-1启动子序列-636^+17、-449^+17-、276^+17 bp 3个片段时,荧光素酶活性显著增高;共转染过氧化体增殖物激活型受体α(PPARα)表达质粒(PPAR-αpSG5)的细胞在非诺贝特诱导下PAI-1转录活性显著降低。结论非诺贝特可以抑制HepG2细胞PAI-1mRNA表达及其活性,调节PAI-1的基因转录,PPARα参与非诺贝特对PAI-1基因的表达调控。
文摘目的探讨肾母细胞瘤基因1(WT1)对鼻咽癌细胞(CNE)-2表皮生长因子受体(EGFR)的靶向作用,证实WT1对CNE-2免疫活性的影响。方法采用WT1对CNE-2进行处理,借助蛋白质免疫印迹法(Western blot)对EGFR的蛋白表达情况进行检测,借助CKK-8观察CNE-2增殖能力情况,借助流式细胞术观察CNE-2凋亡情况,借助基质胶侵袭试验(Transwell)测定CNE-2侵袭情况。结果 Western blot测定结果显示,采用WT1组信号强度明显低于二甲亚砜(DMSO)组和空白组,差异均有统计学意义(P<0.05);WT1组检测灰度值为0.358±0.148,DMSO组为1.370±0.068,空白组为1.390±0.077,WT1组检测灰度值明显低于DMSO组和空白组,差异均有统计学意义(P<0.05)。CKK-8检测结果显示,采用WT1进行处理后3、6、9、12h,抑制值表现为逐渐增大,差异有统计学意义(P<0.05)。流式细胞术检测结果显示,空白组细胞凋亡率为(0.832±0.207)%,DMSO组为(2.081±1.221)%,WT1组为(20.731±1.104)%,WT1组细胞凋亡率高于空白组和DMSO组,差异均有统计学意义(P<0.05)。Transwell测定结果显示,DMSO组穿透细胞数量为67.21±2.17,WT1组为20.13±2.25,WT1组穿透细胞数量低于DMSO组,差异有统计学意义(P<0.05)。结论 WT1抗原能够作为CNE-2EGFR的靶向基因,可较好地实现对CNE-2EGFR免疫活性的抑制。
文摘鸡核受体辅激活蛋白1基因(NCOA1)mRNA的表达对性腺的发育、成熟均起关键作用,并且与繁殖性状相关。为了建立荧光定量PCR技术检测鸡NCOA1表达量的方法,根据GenBank中NCOA1基因序列设计合成了引物,以β-action基因为内参基因,对荧光定量PCR方法进行了评估。结果表明:构建的标准曲线线性关系良好,建立的NCOA1基因荧光定量PCR检测方法灵敏度高、特异性强,准确可靠。建立的SYBR Green I实时荧光定量PCR方法可以测定鸡NCOA1mRNA表达水平的相对含量。
