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Since "normal" carboxysomes represented the majority of microcompartments purified from the mutant by a wide margin, these results suggested that, despite their apparently icosahedral shape and close-to-wild type protein composition, mutant carboxysomes lacking CsoS4A and CsoS4B were more permeable to Ci than wild type particles.
Indeed, we found that the secondary response was significantly reduced by the Ca2+-ATPase inhibitor, thapsigargin, which depletes IP3 sensitive stores of Ca2+.The delayed rise in Ca2+ was also found to be reduced in the dnc mutant by a value that is in the same range of PDE reduction that is reported for the dnc1 mutant [31].
We should stress here that JH.D is not a knock-out strain; instead, mutants were constructed by knock-in of a cbfA genomic DNA fragment containing a premature amber stop codon that is partially suppressed in the mutant by a co-transformed amber suppressor tRNA gene [8].
To evaluate the contribution of low-pH-stable N1 NA to virus replication, we generated reverse genetics viruses (in the A/WSN/33 H1N1 background) possessing the low-pH-unstable wild-type USSR77 NA (USSR77NA) or the low-pH-stable N1 NA (mutant) by a deletion of Thr at 435 and 2 alterations from Arg to Gln at 430 and from Asn to Gly at 455 of the USSR77 NA (Figure 5A).
Wan et al. defined a high-activity mutant by a basal BRAF kinase activity higher than that of oncogenic RAS-activated wild-type BRAF, an intermediate-activity mutant by a basal BRAF kinase activity between those of wild-type BRAF and oncogenic RAS-activated wild-type BRAF, and a kinase-impaired mutant by a basal BRAF kinase activity lower than that of wild-type BRAF [ 13].
Quantitative RT PCR determined that expression of Dsp and Calb1 decreases in the mutant by a factor of 14.7 (P<0.005) and 4.8 (P<0.05), respectively.
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We recently reported a sex mutant by an iodine staining-based screening of a rice mutant collection (Hirose et al. [2013]).
We have developed a powerful method for screening a library of GA mutants by a combination of GA display and SBD mutagenesis on the yeast-cell surface.
In addition, the base-line frequency of micronucleated erythrocytes (MN-RETs) by flow cytometry, HPRT mutants by a lymphocyte cloning assay, and chromosome aberrations by FISH painting were determined from peripheral blood samples.
We next measured the growth suppression activity of RB mutants by a short-term assay of BrdU-incorporation and a long-term assay of flat cell formation.
It is noteworthy that catalytic activity can be rescued in some of the HDAC8 mutants by a small-molecule activator.
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