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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.
In the five instances where fitness was measured for both isogenic ΔflaA and ΔmotAB mutants, there were statistical differences between the non-motile strains in fitness only for the RM2387 and RM2992 backgrounds on broccoli with the ΔflaA mutants reduced in comparison with the ΔmotAB mutants by a factor of five.
It is noteworthy that catalytic activity can be rescued in some of the HDAC8 mutants by a small-molecule activator.
Bithiazoles, however, appear to rescue CFTR processing mutants by a different mechanism because bithiazoles have an additive effect on maturation when used in combination with VX-809.
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Consequently, we could alleviate the growth phenotype and still retain the low xylan levels of the irx mutants by an engineering approach that enables the re-introduction of xylan biosynthesis specifically in the xylem vessels of xylan deficient irx mutants.
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.
The mammalian phocein has been linked to endocytosis (Baillat et al. 2002), therefore we tested endocytosis in the ∆Smmob3 mutant by a FM4-64 uptassayssassay
This model implies that wild-type p53 acts as a 'caretaker' to maintain genetic uniformity in cell populations, and that p53 inactivation increases the probability of occurrence of a viable cellular mutant by a factor of about ten.
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