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To identify the mechanism of action of CIN, we examined the effects of CIN on life span in three mutants implicated in major longevity pathways.
Arbitrary PCR and DNA sequencing analysis revealed that in three mutants the transposon was inserted in the coding regions of exsA and vfr, respectively, which are known T3SS regulatory genes [26], [31].
Although we saw Lefty1 in only a single wild type embryo, possibly due to the transient nature of expression, we nonetheless observed expression in three mutants, whereas Dll1 null mutants have no Lefty1.
In most cases, SlCaMs in three mutants had higher expression than those in wild-type.
Lutein was not altered in LL and only slightly higher in three mutants in HL conditions compared to the wild-type strain.
Southern hybridizations confirmed single insertions at the expected site for most thermotargetrons, but additional off-target insertions were detected in three mutants (CipA-ΔXDocII, CipA-ΔCBM-2, and ΔSdbA) (see Additional file 3).
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An electrophoretic mobility-shift assay and Ku antisera were used to show that Ku or a closely related protein was deficient in three mutant hamster cell lines from x-ray-sensitive complementation group 5, which is characterized by defects in DNA double-strand break repair and V(D J recombination.
Initial characterization revealed peri- and postnatal lethal phenotypes in three mutant strains (Fendrr, Peril, and Mdgt), the latter two exhibiting incomplete penetrance and growth defects in survivors.
Furthermore, we determined whether the tests could be used to monitor the onset and progression of motor dysfunction in three mutant strains with distinct aetiologies.
Fig. 3 shows examples of wheel running activity during the first hour of the night in three mutant mouse strains with previously recorded motor deficits.
Rational enzyme engineering approach resulted in four mutants: G238N, E232A, D103V and Q112H.
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