A combination of assays reveals biomass differences in biofilms formed by Escherichia coli mutants.
A combination of assays reveals biomass differences in biofilms formed by Escherichia coli mutants.
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综合分析揭示了大肠杆菌突变体形成的生物膜的生物量差异。
DOI：
10.1111/j.1472-765x.2009.02659.x
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发表时间：
2009
影响因子：
2.4
通讯作者：
Pruss,BM
中科院分区：
文献类型：
--
作者：
Sule,P;Wadhawan,T;Carr,NJ;Horne,SM;Wolfe,AJ;Pruss,BM
目的：本研究的目的是开发一种测定系统，该系统可以量化由大肠杆菌K-12菌株的不同同基因突变体形成的生物膜中的生物量。方法和结果：所报道的测定基于来自Promega的BacTiter-Glo ™测定，使用生物发光来检测细胞内ATP浓度，其与活细菌细胞数相关。将用该ATP测定法获得的定量数据与用常规结晶紫测定法获得的定量数据进行比较。作为一个定性控制，扫描电子显微镜performed.Conclusions：ATP测定，结晶紫测定和扫描电子显微镜得到了类似的结果为6个测试的8株。对于剩下的两个菌株，从扫描电子显微镜的图像证实了结果从ATP assay.Significance和影响的研究：ATP测定，与其他定量和定性分析相结合，将使我们能够进行遗传研究的监管网络的基础上的早期步骤inE。大肠杆菌生物膜形成。
Aims:The aim of this study was to develop an assay system that can quantify the amount of biomass in biofilms formed by different isogenic mutants of anEscherichia coliK‐12 strain.Methods and Results:The reported assay, which is based on the BacTiter‐Glo™ assay from Promega, uses bioluminescence to detect the intracellular concentration of ATP, which correlates with viable bacterial cell numbers. The quantitative data obtained with this ATP assay were compared to those obtained with the conventional crystal violet assay. As a qualitative control, scanning electron microscopy was performed.Conclusions:The ATP assay, the crystal violet assay and scanning electron microscopy yielded similar results for six of the eight strains tested. For the remaining two strains, the images from the scanning electron microscopy confirmed the results from the ATP assay.Significance and Impact of the Study:The ATP assay, in combination with other quantitative and qualitative assays, will allow us to perform genetic studies on the regulatory network that underlies the early steps inE. colibiofilm formation.