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purified recombinant enzyme, wild-type or selenomethionine-labeled, 10-15 mg/ml protein, hanging drop vapour diffusion method, from 0.1 M MES, pH 6.5, and 3-10% PEG 20000, microseeding, 20Â°C, cryoprotection by 25% v/v glycerol, X-ray diffraction structure determination and analysis at 1.55-1.70 A resolution, modeling
the enzyme mutant K23 of Brucella abortus fails to replicate in mouse macrophages and HeLa cells, and shows less virulence than the wild type in mice, Under nitric oxide (NO) stress, the growth of the DAP mutant in vitro decreases, and it also has less capability to reduce NO than the wild type, intracellular replication of the DAP mutant is partially restored by pretreatment of macrophages with the NO synthase inhibitor 1-phenyl-imidazole and the level of expression of the NO reductase gene norB in the DAP mutant is lower than that in the wild type
gene pbp4B, DNA and amino acid sequence determination and analysis, the pbp4B gene is located at the zipA-dapA region, subcloning in strain DH5alpha, expression of N-terminally His-tagged enzyme in strain BL21(DE3) membranes