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Background
The sexually transmitted disease, gonorrhea, is a serious health problem in developed as well as in developing countries, for which treatment continues to be a challenge. The recent completion of the genome sequence of the causative agent, Neisseria gonorrhoeae, opens up an entirely new set of approaches for studying this organism and the diseases it causes. Here, we describe the initial phases of the construction of an expression-capable clone set representing the protein-coding ORFs of the gonococcal genome using a recombination-based cloning system.Results
The clone set thus far includes 1672 of the 2250 predicted ORFs of the N. gonorrhoeae genome, of which 1393 (83%) are sequence-validated. Included in this set are 48 of the 61 ORFs of the gonococcal genetic island of strain MS11, not present in the sequenced genome of strain FA1090. L-arabinose-inducible glutathione-S-transferase (GST)-fusions were constructed from random clones and each was shown to express a fusion protein of the predicted size following induction, demonstrating the use of the recombination cloning system. PCR amplicons of each ORF used in the cloning reactions were spotted onto glass slides to produce DNA microarrays representing 2035 genes of the gonococcal genome. Pilot experiments indicate that these arrays are suitable for the analysis of global gene expression in gonococci.Conclusion
This archived set of Gateway® entry clones will facilitate high-throughput genomic and proteomic studies of gonococcal genes using a variety of expression and analysis systems. In addition, the DNA arrays produced will allow us to generate gene expression profiles of gonococci grown in a wide variety of conditions. Together, the resources produced in this work will facilitate experiments to dissect the molecular mechanisms of gonococcal pathogenesis on a global scale, and ultimately lead to the determination of the functions of unknown genes in the genome. 相似文献23.
Glasner JD Yang CH Reverchon S Hugouvieux-Cotte-Pattat N Condemine G Bohin JP Van Gijsegem F Yang S Franza T Expert D Plunkett G San Francisco MJ Charkowski AO Py B Bell K Rauscher L Rodriguez-Palenzuela P Toussaint A Holeva MC He SY Douet V Boccara M Blanco C Toth I Anderson BD Biehl BS Mau B Flynn SM Barras F Lindeberg M Birch PR Tsuyumu S Shi X Hibbing M Yap MN Carpentier M Dassa E Umehara M Kim JF Rusch M Soni P Mayhew GF Fouts DE Gill SR Blattner FR Keen NT Perna NT 《Journal of bacteriology》2011,193(8):2076-2077
Dickeya dadantii is a plant-pathogenic enterobacterium responsible for the soft rot disease of many plants of economic importance. We present here the sequence of strain 3937, a strain widely used as a model system for research on the molecular biology and pathogenicity of this group of bacteria. 相似文献
24.
Aïda Boughammoura Thierry Franza Alia Dellagi Camille Roux Berthold Matzanke-Markstein Dominique Expert 《Biometals》2007,20(3-4):347-353
The enterobacterial pathogen Erwinia chrysanthemi causes soft rot diseases on a wide range of plants, including the model plant Arabidopsis thaliana. This bacterium proliferates in the host by secreting a set of pectin degrading enzymes responsible for symptom development.
In addition, survival of this bacterium in planta requires two high-affinity iron acquisition systems mediated by siderophores and protective systems against oxidative damages,
suggesting the implication by both partners of accurate mechanisms controlling their iron homeostasis under conditions of
infection. In this review, we address this question and we show that ferritins both from the pathogen and the host are subtly
implicated in the control of this interplay. 相似文献
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Multi-site phosphorylation of the protein tyrosine phosphatase, PTP1B: identification of cell cycle regulated and phorbol ester stimulated sites of phosphorylation. 总被引:16,自引:4,他引:12 下载免费PDF全文
The non-transmembrane protein tyrosine phosphatase, PTP1B, comprises 435 amino acids, of which the C-terminal 114 residues have been implicated in controlling both localization and function of this enzyme. Inspection of the sequence of the C-terminal segment reveals a number of potential sites of phosphorylation. We show that PTP1B is phosphorylated on seryl residues in vivo. Increased phosphorylation of PTP1B is seen to accompany the transition from G2 to M phase of the cell cycle. Two major tryptic phosphopeptides appear in two-dimensional maps of PTP1B from mitotic cells. One of these comigrates with the peptide generated following phosphorylation of PTP1B in vitro at Ser386 by the mitotic protein Ser/Thr kinase p34cdc2:cyclin B. The site of phosphorylation that is responsible for the pronounced retardation in the electrophoretic mobility of PTP1B from mitotic cells has been identified by site directed mutagenesis as Ser352. The identify of the kinase responsible for this modification is presently unknown. We also show that stimulation of HeLa cells with the phorbol ester TPA enhances phosphorylation of PTP1B. Two dimensional phosphopeptide mapping reveals that the bulk of the phosphate is in a single tryptic peptide. The site, identified as Ser378, is also the site of phosphorylation by protein kinase C (PKC) in vitro. Thus the TPA-stimulated phosphorylation of PTP1B in vivo appears to result directly from phosphorylation by PKC. The effect of phosphorylation on the activity of PTP1B has been examined in immunoprecipitates from TPA-treated and nocodazole-arrested cells. TPA treatment does not appear to affect activity directly, whereas the activity of PTP1B from nocodazole-arrested cells is only 70% of that from asynchronous populations. 相似文献
27.
We examine the competitive implications of a firm's ability to change over its facility for the manufacture of successive generations of high-volume products with short life cycles. This ability is known as changeover flexibility. The model introduced extends the existing literature in several directions. First, the model offers explicit treatment of the critical relationships between market entry time, changeover flexibility, product life cycles, and profit. Second, the model explicitly considers the effect of early market entry on the accumulation of manufacturing experience (learning), which reduces the unit production cost. Third, the product's optimal selling price is determined and its relation to the firm's changeover flexibility is examined. Last, facility flexibility is permitted to vary over a continuum. Therefore, we are able to capture decision making concerning the optimal degree of changeover flexibility. Both analytic and numerical results are reported, demonstrating the link between the operations and marketing domains in the context of a firm's optimal entrance and exit strategies. Among the key findings are (1) a firm more capable of reducing operating costs through learning over short life cycles optimally invests in more changeover flexibility, charges higher prices, and obtains greater profit; and (2) as the cost of flexible technologies decrease, a firm optimally increases its investment in changeover flexibility, enters markets earlier, and charges higher average prices over the product's life cycle. 相似文献
28.
Analysis of the Erwinia chrysanthemi ferrichrysobactin receptor gene: resemblance to the Escherichia coli fepA-fes bidirectional promoter region and homology with hydroxamate receptors. 下载免费PDF全文
The fct cbsCEBA operon from the Erwinia chrysanthemi 3937 chrysobactin-dependent iron assimilation system codes for transport and biosynthetic functions. The sequence of the fct outer membrane receptor gene was determined. The fct promoter region displays a strong resemblance to the Escherichia coli bidirectional intercistronic region controlling the expression of the fepA-entD and fes-entF operons. An apparent Fur-binding site was shown to confer iron regulation on an fct::lac fusion expressed on a low-copy-number plasmid in a Fur-proficient E. coli strain. The fct gene consists of an open reading frame encoding a 735-amino-acid polypeptide with a signal sequence of 38 residues. The Fct protein has 36% sequence homology with the E. coli ferrichrome receptor FhuA and the Yersinia enterocolitica ferrioxamine receptor FoxA. On the basis of secondary-structure predictions and these homologies, we propose a two-dimensional folding model for Fct. 相似文献
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A 60 kd cdc2-associated polypeptide complexes with the E1A proteins in adenovirus-infected cells 总被引:60,自引:0,他引:60
p60 is a cellular protein that binds to the adenovirus E1A protein complex in virally infected or transformed human cells. In both infected and uninfected cells, p60 was found in a complex with the cdc2 protein kinase. Immune complexes containing p60 and cdc2 display a cell cycle-dependent histone H1 kinase activity that is most active in interphase. The previously described cdc2-p62/cyclin complex also acts as a histone H1 kinase but is maximally active in mitotic metaphase. The shift in the timing of activation of different cdc2-containing complexes suggests that each might play a distinct role in regulation of the cell cycle. 相似文献