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101.
Kemin Tan Gekleng Chhor T. Andrew Binkowski Robert P. Jedrzejczak Magdalena Makowska-Grzyska Andrzej Joachimiak 《The Journal of biological chemistry》2014,289(18):12232-12244
The overproduction of polysaccharide alginate is responsible for the formation of mucus in the lungs of cystic fibrosis patients. Histidine kinase KinB of the KinB-AlgB two-component system in Pseudomonas aeruginosa acts as a negative regulator of alginate biosynthesis. The modular architecture of KinB is similar to other histidine kinases. However, its periplasmic signal sensor domain is unique and is found only in the Pseudomonas genus. Here, we present the first crystal structures of the KinB sensor domain. The domain is a dimer in solution, and in the crystal it shows an atypical dimer of a helix-swapped four-helix bundle. A positively charged cavity is formed on the dimer interface and involves several strictly conserved residues, including Arg-60. A phosphate anion is bound asymmetrically in one of the structures. In silico docking identified several monophosphorylated sugars, including β-d-fructose 6-phosphate and β-d-mannose 6-phosphate, a precursor and an intermediate of alginate synthesis, respectively, as potential KinB ligands. Ligand binding was confirmed experimentally. Conformational transition from a symmetric to an asymmetric structure and decreasing dimer stability caused by ligand binding may be a part of the signal transduction mechanism of the KinB-AlgB two-component system. 相似文献
102.
Motomi Ito Kuniaki Watanabe Yoko Kita Takayuki Kawahara DJ Crawford Tetsukazu Yahara 《Journal of plant research》2000,113(1):79-89
Eupatorium were examined by sequencing the internal transcribed spacers (ITS) of nuclear ribosomal DNA and restriction site analysis
of chloroplast DNA. Molecular data provided strong evidence that (1) this genus originated in North America, (2) the genus
diverged into three morphological species groups, Eutrochium, Traganthes and Uncasia in North America, and (3) one of the North American Uncasia lineages migrated into temperate Europe and eastern Asia over the Bering land bridge. The estimated divergence times support
a late Miocene to early Pliocene migration from North America to Eurasia via the Bering land bridge. A European species was
sister to all of the eastern Asian species examined. The disjunct distribution pattern of the genus Eupatorium is incongruent with the classical Arcto-Tertiary geoflora concept.
Received 13 September 1999/ Accepted in revised form 4 January 2000 相似文献
103.
Although efficient methods exist to assemble synthetic oligonucleotides into genes and genomes, these suffer from the presence of 1–3 random errors/kb of DNA. Here, we introduce a new method termed consensus shuffling and demonstrate its use to significantly reduce random errors in synthetic DNA. In this method, errors are revealed as mismatches by re-hybridization of the population. The DNA is fragmented, and mismatched fragments are removed upon binding to an immobilized mismatch binding protein (MutS). PCR assembly of the remaining fragments yields a new population of full-length sequences enriched for the consensus sequence of the input population. We show that two iterations of consensus shuffling improved a population of synthetic green fluorescent protein (GFPuv) clones from ~60 to >90% fluorescent, and decreased errors 3.5- to 4.3-fold to final values of ~1 error per 3500 bp. In addition, two iterations of consensus shuffling corrected a population of GFPuv clones where all members were non-functional, to a population where 82% of clones were fluorescent. Consensus shuffling should facilitate the rapid and accurate synthesis of long DNA sequences. 相似文献
104.
105.
Several studies have indicated that olfactory responses are impeded by
amiloride. Therefore, it was of interest to see whether, and if so which,
olfactory epithelial cellular compartments have amiloride- sensitive
structures. Using ultrastructural methods that involved rapid freezing,
freeze-substitution and low temperature embedding of olfactory epithelia,
this study shows that, in the rat, this tissue is immunoreactive to
antibodies against amiloride sensitive Na(+)- channels. However, microvilli
of olfactory supporting cells, as opposed to receptor cilia, contained most
of the immunoreactive sites. Apices from which the microvilli sprout and
receptor cell dendritic knobs had much less if any of the
amiloride-antibody binding sites. Using a direct ligand-binding
cytochemical method, this study also confirms earlier ones that showed that
olfactory receptor cell cilia have Na+, K(+)-ATPase. It is proposed that
supporting cell microvilli and the receptor cilia themselves have
mechanisms, different but likely complementary, that participate in
regulating the salt concentration around the receptor cell cilia. In this
way, both structures help to provide the ambient mucous environment for
receptor cells to function properly. This regulation of the salt
concentration of an ambient fluid environment is a function that the
olfactory epithelium shares with cells of transporting epithelia, such as
those of kidney.
相似文献
106.
Transgenic Research - 相似文献
107.
Spawning behavior of lake sturgeon (Acipenser fulvescens) 总被引:1,自引:1,他引:0
108.
109.
W. S. Ramsey W. Hertl E. D. Nowlan N. J. Binkowski 《In vitro cellular & developmental biology. Plant》1984,20(10):802-808
Summary Photoelectron spectroscopic examination of treated plastic surfaces showed that surface oxidation, primarily as carboxyl groups,
was responsible for formation of good growth surfaces. Gas-plasma studies indicated that only very short exposures were required
and that the effect was confined to a thin surface layer that produced adhesive surfaces. Highly adhesive surfaces were produced
using oxidizing chemicals. Studies with a polymeric ester demonstrated the importance of unesterified carboxyl groups for
high adhesiveness. 相似文献