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Figure 6
In vitro ribonuclease-activity assay. (a) Fluorescence measurements as a function of time during the addition of increasing amounts of MazF. Various concentrations of MazF monomer were prepared; at concentrations greater than 4 µM, the reaction was saturated. (b) A metal-dependent ribonuclease-activity assay was conducted using several divalent metal ions. Notable changes were not observed in this assay. (c) An in vitro ribonuclease assay of K. pneumoniae MazF. The RFU obtained with 4 µM WT MazF was taken as 100%. (d) An in vivo cell-growth assay of K. pneumoniae MazF. The growth of E. coli Rosetta (DE3) pLysS was monitored over 6 h after induction of WT MazF and several mutants of MazF. The error bars represent the standard error of the means from three independent experiments.

IUCrJ
Volume 8| Part 3| May 2021| Pages 362-371
ISSN: 2052-2525
https://doi.org/10.1107/S2052252521000452