Графический дизайн как визуальный язык межкультурного взаимодействия

This article describes how visual graphics language as a sign system can be in contact with the audience, overcoming the language barrier. In terms of graphic design it can be available to transfer information, and even affect the viewer, causing artistic and emotional reflection.Эта статья о том, как визуальный язык графики в виде знаковой символики может входить в контакт со зрителем, преодолевая языковый барьер. На языке графического дизайна можно доступно передать информацию и даже воздействовать на зрителя, вызывая при этом художественно-эмоциональные образы

Relationships between SAV576 and SAV577.

<p>(<b>A</b>) Comparison of DNA-binding affinity of SAV576 and SAV577 with probes 1 and 577p. Each lane contained 0.15 nM labeled probe. White arrows: free probes. Black arrows: DNA-protein complexes. (<b>B</b>) Competitive EMSAs of probe 1 with His₆-SAV576 and His₆-SAV577 proteins. End-labeled probe 1 was incubated with the indicated concentrations of His₆-SAV576 or His₆-SAV577. Lanes 1 and 1′: control reactions (no protein added). Diagonal arrows: two complexes in lanes 3 and 3′. (<b>C</b>) GST pull-down assays of SAV576 and SAV577 from <i>E. coli</i> whole cell lysate. GST- and His₆-tagged proteins were co-expressed in <i>E. coli</i>, lysed by sonication, and subjected to GST pull-down assay and Western blotting using anti-GST and anti-His antibodies, respectively. Lanes 1, cell lysate before IPTG induction. Lanes 2, cell lysate after induction. Lanes 3, GST pull-down.</p

Identification of <i>SAV577</i> as a target gene of SAV576.

<p>(<b>A</b>) Real-time RT-PCR analysis of <i>SAV577</i> transcription levels for ATCC31267 (WT) and <i>SAV576</i> deletion mutant D576 grown in FM-II on days 2 and 6. Relative values were obtained using <i>hrdB</i> as a reference. Error bars, standard deviations (<i>n</i> = 3). ***<i>P</i><0.001 as determined by Student's <i>t</i>-test. (<b>B</b>) ChIP assays. Anti-SAV576 antibody was used to immunoprecipitate SAV576-DNA complexes from ATCC31267 and D576 cells treated with formaldehyde. The DNAs used for PCR were total DNA prior to immunoprecipitation (positive control: lanes “+”), immunoprecipitated DNA with anti-SAV576 antibody (experimental sample: lanes “S”), and immunoprecipitated DNA with rabbit preimmune serum (negative control: lanes “IgG”). The <i>hrdB</i> promoter region was used as a control. (<b>C</b>) EMSAs of the interaction of probe 577p with purified His₆-SAV576 protein. Each lane contained 0.3 nM labeled probe. An approximately 100-fold excess of the unlabeled probe was used in competitive assays. BSA was used as a negative control for SAV576 protein. Labeled 262-bp <i>hrdB</i> promoter region (probe <i>hrdBp</i>) was used to eliminate non-specific binding of the SAV576 protein. The free probe is indicated by solid arrow, and the retarded DNA fragment is indicated by parentheses.</p

SAV577 directly represses <i>SAV575</i> and <i>SAV576</i> expression.

<p>(<b>A</b>) Effect of <i>SAV577</i> deletion on expression of <i>aveR</i>, <i>aveA1</i>, <i>SAV575</i>, <i>SAV576</i> and <i>SAV577</i>, and detection of <i>SAV575</i> transcription level in various <i>S. avermitilis</i> strains. <i>SAV577′</i>, 136-bp transcript from <i>SAV577</i> promoter region and the remainder ORF in D577. Error bars, standard deviations (<i>n</i> = 3). ***<i>P</i><0.001 as determined by Student's <i>t</i>-test. ns, not significant. (<b>B</b>) Schematic representation of the relative positions of probes used for EMSAs. The lengths and positions of probes 1–7 were described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099224#pone.0099224-Guo1" target="_blank">[20]</a>. Probe 577p, a 209-bp DNA fragment from positions −159 to −367 relative to the <i>SAV577</i> start codon, covering the putative transcriptional start point of <i>SAV577</i>. (<b>C</b>) EMSAs of the interaction of probes with purified His₆-SAV577 protein. Each lane contained 0.3 nM labeled probe. (<b>D</b> and <b>E</b>) DNase I footprinting assay of SAV577 on the <i>SAV575</i> (<b>D</b>) and <i>SAV576</i> (<b>E</b>) promoter regions. The fluorograms represent control DNA (10 µM BSA) and protection reactions with increasing concentrations (0.08, 0.4, 0.8 µM) of His₆-SAV577 protein.</p

The possible regulatory pathway of SAV576/SAV577/SAV575 on avermectin biosynthesis in <i>S. avermitilis</i>.

<p>Two similar TFRs SAV576 and SAV577 both repress avermectin biosynthesis indirectly by (i) repressing <i>SAV575</i> expression directly, and SAV575 may provide precursors for avermectin biosynthesis; (ii) controlling unknown regulatory gene(s) that directly (or indirectly) regulate expression of the pathway-specific activator gene <i>aveR</i>, and AveR activates avermectin biosynthesis by activating the transcription of biosynthetic structural genes. SAV576 and SAV577 repress each other's expression. However, SAV576 is autoregulated whereas SAV577 is not. Arrows indicate activation, and bars indicate repression. Solid lines indicate direct control, and dotted lines indicate unknown routes.</p

Two Adjacent and Similar TetR Family Transcriptional Regulator Genes, <i>SAV577</i> and <i>SAV576</i>, Co-Regulate Avermectin Production in <i>Streptomyces avermitilis</i>

<div><p><i>Streptomyces avermitilis</i> is an important bacterial species used for industrial production of avermectins, a family of broad-spectrum anthelmintic agents. We previously identified the protein SAV576, a TetR family transcriptional regulator (TFR), as a downregulator of avermectin biosynthesis that acts by controlling transcription of its major target gene <i>SAV575</i> (which encodes cytochrome P450/NADPH-ferrihemoprotein reductase) and <i>ave</i> genes. <i>SAV577</i>, another TFR gene, encodes a SAV577 protein that displays high amino acid homology with SAV576. In this study, we examined the effect of SAV577 on avermectin production and the relationships between SAV576 and SAV577. SAV577 downregulated avermectin biosynthesis indirectly, similarly to SAV576. SAV576 and SAV577 both directly repressed <i>SAV575</i> transcription, and reciprocally repressed each other's expression. <i>SAV575</i> transcription levels in various <i>S. avermitilis</i> strains were correlated with avermectin production levels. DNase I footprinting and electrophoretic mobility shift assays indicated that SAV576 and SAV577 compete for the same binding regions, and that DNA-binding affinity of SAV576 is much stronger than that of SAV577. GST pull-down assays revealed no direct interaction between the two proteins. Taken together, these findings suggest that SAV577 regulates avermectin production in <i>S. avermitilis</i> by a mechanism similar to that of SAV576, and that the role of SAV576 is dominant over that of SAV577. This is the first report of two adjacent and similar TFR genes that co-regulate antibiotic production in <i>Streptomyces.</i></p></div

Avermectin production and growth of wild-type strain ATCC31267 and <i>SAV577</i> mutant strains.

<p>(<b>A</b>) Comparison of avermectin production in various <i>S. avermitilis</i> strains grown in FM-I medium for 10 days. WT, wild-type strain ATCC31267. WT/pKC1139, ATCC31267 carrying control plasmid pKC1139. D576, <i>SAV576</i> deletion mutant. D577, <i>SAV577</i> deletion mutant. D577/pSET152-577, complementation strain of D577. WT/pKC1139-577, <i>SAV577</i> overexpression strain. D576-577, <i>SAV576-SAV577</i> double deletion mutant. (<b>B</b> and <b>C</b>) Effect of <i>SAV577</i> deletion on avermectin production (<b>B</b>) and growth (<b>C</b>) of <i>S. avermitilis</i> grown in soluble medium FM-II. Solid squares, ATCC31267; Solid circles, D577. Results are shown as mean ± SD for three independent experiments. **<i>P</i><0.01 and ***<i>P</i><0.001 as determined by Student's <i>t</i>-test. ns, not significant.</p

MOESM4 of Efficient heterologous expression of an alkaline lipase and its application in hydrolytic production of free astaxanthin

Additional file 4: Table S2. Effects of acetone concentration on Lipase-YH extraction

MOESM1 of Efficient heterologous expression of an alkaline lipase and its application in hydrolytic production of free astaxanthin

Additional file 1: Fig. S1. Analysis of Penicillium cyclopium var. albus lipase gene. A: Signal peptide analysis of the gene by signalP-4.0 Server program. B: Signal peptide and propeptide of the gene. A 20-a.a. signal peptide and 7-a.a. propeptide were found at the start of the 258-a.a. mature lipase

Data_Sheet_1_Organic Peroxide-Sensing Repressor OhrR Regulates Organic Hydroperoxide Stress Resistance and Avermectin Production in Streptomyces avermitilis.PDF

<p>The bacterium Streptomyces avermitilis is an industrial-scale producer of avermectins, which are important anthelmintic agents widely used in agriculture, veterinary medicine, and human medicine. During the avermectin fermentation process, S. avermitilis is exposed to organic peroxides generated by aerobic respiration. We investigated the role of MarR-family transcriptional regulator OhrR in oxidative stress response and avermectin production in S. avermitilis. The S. avermitilis genome encodes two organic hydroperoxide resistance proteins: OhrB1 and OhrB2. OhrB2 is the major resistance protein in organic peroxide stress responses. In the absence of organic peroxide, the reduced form of OhrR represses the expression of ohrB2 gene by binding to the OhrR box in the promoter region. In the presence of organic peroxide, the oxidized form of OhrR dissociates from the OhrR box and the expression of ohrB2 is increased by derepression. OhrR also acts as a repressor to regulate its own expression. An ohrR-deletion mutant (termed DohrR) displayed enhanced avermectin production. Our findings demonstrate that OhrR in S. avermitilis represses avermectin production by regulating the expression of pathway-specific regulatory gene aveR. OhrR also plays a regulatory role in glycolysis and the pentose phosphate (PP) pathway by negatively controlling the expression of pykA2 and ctaB/tkt2-tal2-zwf2-opcA2-pgl.</p