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. Author manuscript; available in PMC: 2009 May 29.
Published in final edited form as: Channels (Austin). 2008 Nov 27;2(6):413–418. doi: 10.4161/chan.2.6.6874

Figure 5.

Figure 5

Inactivation mutations and sensitivity of the assay. (A) The inactivation mutation E71A fails to complement growth of TK2420 in 7.5 mM K+ K0 media indicating that removal of inactivation in the selectivity filter gate is not enough to open the channel in vivo. The double mutant H25L/E71A complements growth well demonstrating that the E71A mutation does not fail to complement due to ‘toxicity’. (B) Mutant and wild-type KcsA were grown in high K+ K0 media (60 mM) and expression was induced as indicated by the arrow. Wild-type KcsA, E71A and A108T exhibited robust growth over this 8 hour period. By contrast A108D cultures started to die off. Consequently such overactive mutations would not be found in this gain-of-function screen due to their intrinsic toxicity. Data shown are mean ± S.D., n = 3.