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Jimena Rinaldi Ignacio Fernández Lucía M. Poth William E. Shepard Martin Savko Fernando A. Goldbaum Sebastián Klinke 《Biochemistry and Biophysics Reports》2018
The pathogenic bacterium Brucella abortus codes for a multi-domain dimeric cytoplasmic histidine kinase called LOV-HK, which is a key blue light-activated virulence factor in this microorganism. The structural basis of the light activation mechanism of this protein remains unclear. In this work, full-length LOV-HK was cloned, expressed and purified. The protein was activated by light and crystallized under a controlled illumination environment. The merge of 14 individual native data sets collected on a single crystal resulted in a complete X-ray diffraction data set to a resolution of 3.70 Å with over 2 million reflections. Crystals belong to space group P212121, with unit-cell parameters a = 95.96, b = 105.30, c = 164.49 Å with a dimer in the asymmetric unit. Molecular replacement with Phaser using the individual domains as search models allowed for the reconstruction of almost the whole protein. Very recently, improved LOV-HK crystals led to a 3.25-Å resolution dataset. Refinement and model building is underway. This crystal model will represent one of the very few examples of a multi-domain histidine kinase with known structure. 相似文献
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ABSTRACTFew regional faunistic treatments of aquatic Heteroptera exist for Mesoamerica. Records given here are the result of three extensive, coincident projects, each generating many specimens of Naucoridae. Thus, presented here is an annotated list of Naucoridae occurring in Belize, with an illustrated taxonomic key and notes on habitat. Three species are new country records, and ten species in four genera are now known from Belize. 相似文献
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Magnetic bead capture of cDNAs from double-stranded plasmid cDNA libraries. 总被引:2,自引:0,他引:2
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We have developed a cDNA library screening method which allows the simultaneous screening of >10 ( 12 ) double-stranded plasmid cDNA molecules with minimal a priori sequence knowledge. A biotinylated, gene-specific oligonucleotide probe along with abutting 'blocking' oligos is hybridized to the plasmid cDNA library and the target plasmid retrieved with paramagnetic streptavidin beads and transformed into Escherichia coli. Multiple rounds of enrichment with a target plasmid represented at 0.002-0.0001% resulted in over one-third positive clones. Our method will be useful for isolating even the rarest cDNAs starting from ESTs, isolated exons or homologous sequence information. 相似文献
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