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Furthermore, using structure-based mutants, we identify a triad of residues in the methyllysine binding pocket that imparts discrimination between monomethyllysine and dimethyllysine.
The mutants we identify here as slow to aggregate (H80R, D101N, D125H, E133ΔE, S134N and V148I) exhibit some of the characteristics of WT SOD1 (see Supplementary Material, Fig. S3).
Despite that many transcription factors are positively affected in the long-lived mutants, we identify only a few negatively affected TF gene set using ChIP-chip banalysislysee (see Table 3), similar to the results by motif enrichment analysis.
Using RNA-Seq analysis of Tfc6p under-expressing mutants, we identify the genome-wide impacts of Pol III complex binding sites, and show that numerous coding and intergenic transcripts were affected in the mutants.
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From a library consisting of 2126 knock-out mutants, we identified 3 mutants with high-flux and 95 mutants with low-flux phenotypes that did not have severe growth defects.
In fact, none of the hypercytotoxic transposon mutants we identified through our screen were in genes required for O-antigen synthesis.
The four mutants we identified as being more cytotoxic to macrophages and phagocytosed via an actin-independent pathway are all normally involved in synthesis of the LPS core.
However, as we discuss in more detail below, there is indirect evidence in other Gram-negative bacteria that mutations that alter the LPS may have phenotypes similar to the four mutants we identified in this manuscript.
Moreover, by employing a variety of protein mutants, we identified the Ig-like domains 4 or 5 of MAG as necessary and sufficient for growth arrest, whereas abolishing MAG's ability to bind to sialic acid did not interfere with its inhibitory activity.
Accordingly, using single gene deletion mutants, we identified yeast genes that control PEA metabolism, including SPO14 (a yeast homologue of the mammalian phospholipase D) that controls PEA production and YJU3 (a yeast homologue of the mammalian monoacylglycerol lipase) that controls PEA inactivation.
Upon screening 200 mutants, we identified three mutants that repeatedly induced less granuloma formation.
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