A high-throughput screen for MscL channel activity and mutational phenotyping.
A novel fluorescence-based screen for bacterial mechanosensitive ion-channel activity has been developed. This assay is capable of clearly distinguishing the previously observed gain of function and loss of function phenotypes for the Escherichia coli mechanosensitive channel of large conductance (Ec-MscL). The method modifies Molecular Probes' Live/Dead BacLight bacterial viability assay to monitor MscL channel activity as a function of bacterial survival from osmotic downshock.
Maurer
J A
JA
Division of Chemistry and Chemical Engineering, California Institute of Technology, Mail Code 164-30 Cr., Pasadena, CA 91125, USA.
Dougherty
D A
DA
D002470:Cell Survival; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D029968:Escherichia coli Proteins; D005453:Fluorescence; D007473:Ion Channels / Q000032:analysis* / Q000235:genetics; D009154:Mutation; D009997:Osmotic Pressure; D010641:Phenotype; D011419:Propidium
D029968:Escherichia coli Proteins; D007473:Ion Channels; C086596:MscL protein, E coli; D011419:Propidium