...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Cytometry: The Journal of the Society for Analytical Cytology >Monitoring of in vitro and in vivo translation of green fluorescent protein and its fusion proteins by fluorescence correlation spectroscopy
【24h】

Monitoring of in vitro and in vivo translation of green fluorescent protein and its fusion proteins by fluorescence correlation spectroscopy

机译：通过荧光相关光谱法监测绿色荧光蛋白及其融合蛋白的体内和体外翻译

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Background: Because the process of protein translation is an event of sparse molecules, the measurement requires high sensitivity. One of the candidates for studying the molecules is fluorescence correlation spectroscopy (FCS), which gleans quantitative information from fluctuating fluorescence signals in a diluted solution. Methods: Using FCS, the translation products of expression plasmid for green fluorescent protein (GFP) and its fusion proteins were measured in vitro and in vivo. Results: In in vitro translation, the number of products increased linearly for 90 min upon concentration of the plasmid. The autocorrelation function for GFP was fitted with a one-component model with a diffusion time of 0.18 ms, which was identical to the value expected from the molecular weight. In the cases of GFP- tagged hypoxia-inducible factor-la and glucocorticoid receptor, each fitting result was significantly improved with a two-component model. The slow component with a diffusion time of G ms appeared to be related to the ribosome or polysome. In response to the addition of dexamethasone, the nuclear translocation from cytosol clearly induced the decrease in number of molecules in the focal point. Conclusions: FCS permits monitoring of the number of molecules translated in vitro and in vivo, the translation rate, and the molecular weight. Cytometry 44:1-6, 2001. (C) 2001 Wiley-Liss, Inc. [References: 25]
机译：背景：由于蛋白质翻译过程是分子稀疏的事件，因此测量需要很高的灵敏度。用于研究分子的候选方法之一是荧光相关光谱（FCS），它可以从稀释溶液中波动的荧光信号中收集定量信息。方法：采用FCS技术，在体外和体内检测绿色荧光蛋白（GFP）表达质粒及其融合蛋白的翻译产物。结果：在体外翻译中，浓缩质粒后90分钟内产物的数量线性增加。 GFP的自相关函数与扩散时间为0.18 ms的单组分模型拟合，该模型与分子量预期的值相同。在GFP标记的缺氧诱导因子-1a和糖皮质激素受体的情况下，采用两组分模型可显着改善每个拟合结果。扩散时间为G ms的慢组分似乎与核糖体或多核糖体有关。响应于地塞米松的添加，来自细胞质的核易位清楚地引起了焦点分子数量的减少。结论：FCS可以监测体外和体内翻译的分子数量，翻译速率和分子量。 Cytometry 44：1-6，2001。（C）2001 Wiley-Liss，Inc. [参考：25]

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号