CaltechAUTHORS
Published October 1, 2001 | Version public
Journal Article Metadata-only

A high-throughput screen for MscL channel activity and mutational phenotyping

Creators

  • 1. ROR icon California Institute of Technology

Abstract

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.

Additional Information

© 2001 Published by Elsevier B.V. Received 8 June 2001, Revised 5 July 2001, Accepted 5 July 2001, Available online 6 September 2001. We are extremely grateful to Ann Batiza and Ching Kung from the University of Wisconsin, Madison for the G22X MscL constructs in pB10b. The MJF465 bacteria strain was a kind gift from Ian Booth of the University of Aberdeen. Donald Elmore, Steve Spronk, Professor Henry Lester, and Professor Douglas Rees provided many helpful discussions. J.A.M. is a recipient of National Institutes of Health Predoctoral Trainee Grant GM-08501. The National Institutes of Health Program Project Grant GM-62532 supported this work.

Additional details

Identifiers

Eprint ID
103010
DOI
10.1016/s0005-2736(01)00390-x
Resolver ID
CaltechAUTHORS:20200505-154609534

Related works

Describes
10.1016/s0005-2736(01)00390-x (DOI)

Funding

NIH Predoctoral Fellowship
GM-08501
NIH
GM-62532

Dates

Created
2020-05-05
Created from EPrint's datestamp field
Updated
2021-11-16
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