Imaging OmpR localization in Escherichia coli
Imaging OmpR localization in Escherichia coli
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DOI：
10.1111/j.1365-2958.2006.05048.x
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发表时间：
2006-03-01
影响因子：
3.6
通讯作者：
Goulian, M
中科院分区：
文献类型：
--
作者：
Batchelor, E;Goulian, M
我们已经使用了一个融合的绿色荧光蛋白的响应调节OmpR的图像的空间分布OmpR在大肠杆菌活细胞。我们观察到增加的OmpR-GFP荧光的病灶，这似乎是由于与组氨酸激酶EnvZ的相互作用。我们还观察到OmpR-GFP与携带OmpR结合位点的质粒簇的共定位，这使我们能够开发一种简单的方法来成像OmpR与活细胞中DNA的结合。我们使用细胞内的峰值荧光强度来量化由于与EnvZ的相互作用或由于结合DNA而导致的OmpR-GFP定位的程度。通过这些试验，我们比较了渗透压和普鲁卡因的影响，认为这两者都能调节EnvZ活性。我们的研究结果表明，至少在我们的生长条件下，普鲁卡因激活EnvZ-OmpR信号，而渗透压，充其量，对EnvZ-OmpR系统的影响很弱。
We have used a fusion of GFP to the response regulator OmpR to image the spatial distribution of OmpR in live cells of Escherichia coli. We observed foci of increased OmpR-GFP fluorescence that appear to be due to interactions with the histidine kinase EnvZ. We also observed colocalization of OmpR-GFP with clusters of plasmids carrying OmpR binding sites, which enabled us to develop a simple method for imaging the binding of OmpR to DNA in live cells. We used the peak fluorescence intensity within cells to quantify the extent of OmpR-GFP localization either due to interactions with EnvZ or due to binding DNA. With these assays we compared the effects of osmolarity and procaine, both of which are believed to modulate EnvZ activity. Our results suggest that, at least under our growth conditions, procaine activates EnvZ-OmpR signalling whereas osmolarity has, at best, a weak effect on the EnvZ-OmpR system.