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Surprisingly, we identified four deletion mutants that improved xylose consumption of laboratory S288C, even in the absence of exogenous XYLA.
To facilitate rapid selection of improved AcrB from the enriched libraries in the absence of a high-throughput method for measuring efflux rate of n-octane, the enriched libraries were preliminarily assayed as variant mixtures to identify the pools with AcrB mutants that improved extrusion of n-octane.
Three known antibiotic substrates of AcrB [ 29] of varying molecular weight – nalidixic acid (232.24 g mol-1), chloramphenicol (323.13 g mol-1) and tetracycline (444.44 g mol-1) – were chosen to investigate the antibiotic resistance of the mutants that improved n-octane efflux (i.e. 1G1, 1G2, N189H, T678S, T678A, Q737L, M844L and M844A).
We have previously used targeted active-site saturation mutagenesis to identify a number of transketolase single mutants that improved activity towards either glycolaldehyde (GA), or the non-natural substrate propionaldehyde (PA).
The genome-wide chemical and genetic screens identified four deletion mutants that improved growth on xylose: RPL20B, ALP1, ISC1, and BUD21.
The rate of α-pinene release was assayed (Table 1) and interestingly, the mutants that improved efflux of n-octane also enhanced the efflux rate of α-pinene (Table 1) despite the structural dissimilarity of the substrates.
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This dataset contains GALR3 point mutants that improve recombinant protein expression and thermal stability of the receptor contained in virus-like particles (VLPs) or obtained by detergent-purification of baculovirus-infected insect cells.
In our experience mutations that improve fluorescent brightness are much more readily identified than mutants that improve photostability.
Maranas and colleagues have previously applied mixed integer linear programming (MILP) to design mutants that improve byproduct yield or identify "minimal reaction sets" that are capable of sustaining a given biomass optimum [ 24, 25].
It may be beneficial to exploit the molecular "barcodes" of yeast mutant collections and microarray-based methods (reviewed in Smith et al. 2010) to identify mutants that improve either recombinant XI/XK or XR/XDH/XK xylose fermentation in anaerobic batch cultures that better replicate industrial-scale biofuel production.
We performed a "chemical suppression" screen by using a previously described collection of temperature-sensitive (TS) mutants (Li et al. 2011) and sought those mutants that showed improved growth in the presence of HU at the otherwise-restrictive temperature where the mutant would normally fail to grow.
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