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Some mutants introduced a small amount of α(1 4) linkages and α(1 2) linkages.
One of the mutants introduced a premature termination codon whereas the other mutation resulted in an amino acid substitution.
Sequencing reverse-transcribed cDNA from mutants and wild-type littermates with oligonucleotide primers for Nrp1 confirmed that a 4 bp insertion in the mutants introduced a premature stop codon at the end of the transmembrane domain and shifted the mRNA reading sequence out of frame, which prevented translation of the cytoplasmic domain (Fig. 1F).
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Our computational protocol serves to design point mutants introducing a conformational shift in the ground-state ensemble.
The R48A/W191F mutant introduced a binding site for N-hydroxyguanidine near the distal heme face and removed the redox active Trp-191 radical site.
High lethality of mutants introduces an error-threshold in a multiplicative fitness landscape.
The hydrophobicity of the wild type and mutant differ because the mutation introduces a very high hydrophobic residue in place of a less hydrophobic residue.
On the contrary, mutation of the second T of the element (T4 in the exon) has no effect on the splicing pattern, whereas mutation of the first T (T3 in the exon) leads to high levels of inclusion only if changed to G. Since the preceding nucleotide in the exon is an A, this mutant introduces an AG just three nucleotides downstream of the natural AG.
Furthermore, the coincidence detection idea is not tested directly, for example with SLAC1 S120A singlesingle and double mutants introduced into the background of a SLAC1 loss-of-function mutant.
Specifically, we generated auxotrophic mutants and introduced a xylose metabolic pathway into the auxotrophic mutants.
To further understand the molecular basis of increased glial phagocytosis in puc mutants, we introduced a reporter for dJNK activity, TRE-eGFP that contains Drosophila AP-1-binding sites fused to the eGFP gene.
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