Transcriptome changes elicited by glucose in GBS.

<p>Comparison of gene expression changes (log2) between mid-log cultures of 2603 V/R wild-type strain and <i>CovRS</i> mutant following challenge with or without glucose. Data for the wild type and mutant strains are shown on the <i>x</i> axis and <i>y</i> axis, respectively.</p

Real-time RT-PCR evaluation of <i>bibA</i> expression in 2603V/R and Δc<i>ovRS</i> strains grown in medium containing 55 mM glucose or in sugar-free medium.

<p>Transcript levels were normalized to the expression level of <i>gyrA</i>. Syber green runs were performed with cDNAs from the same reverse transcription reaction from 1 µg of total RNA. The ΔΔCT method was applied as a comparative method of quantification, using strains grown in sugar free medium as reference. The data are representative of 2 independent experiments, each in triplicate. Error bars, SD.</p

CovR binds to <i>bibA</i> promoter <i>in vivo</i>. (A)

<p>Quantification by qRT-PCR of <i>bibA</i> promoter immunoprecipitated with CovR antiserum in 2603 V/R wild type strain grown in medium devoid of glucose or in the presence of 55mM glucose. <i>cfb</i> promoter and <i>cylX</i> promoter were used as a positive control while <i>sag0017</i> promoter was used as a negative control. The level of PCR products of eluate from the isogenic Δc<i>ovRS</i> deletion mutant grown with or without glucose was negligible. The data are representative of 3 independent experiments, each in triplicate. Error bars, SD. (<b>B)</b> Competitive EMSA experiment. Labelled <i>PbibA</i> fragment (3.3 nM) was incubated without <i>(lane1)</i> or with CovR (2 µM) <i>(lane2</i>–<i>6)</i>, in the presence of different amounts of unlabelled <i>PbibA (lane 3</i>–<i>4)</i>, as a specific competitor, and <i>Psag0017 (lane5</i>–<i>6)</i>, as a non-specific competitor. The labelled DNA was detected by chemioluminescence. <b>(C)</b> CovR phosphorylation increases its affinity for <i>bibA</i> promoter. Electrophoretic mobility shift assay using recombinant CovR (left) and chemically phosphorylated recombinant CovR (right). Labelled <i>PbibA</i> DNA fragment (3.3 nM) was incubated without or with the indicated amounts of CovR. The labelled DNA was detected by chemioluminescence.</p

Differential regulation of gene expression in GBS strain 2603 V/R versus the isogenic Δ<i>CovRS</i> mutant strain after incubation in medium with 55 mM glucose versus a sugars-free complex medium.

<p>White bars indicate the number of glucose-regulated genes in the wild-type strain; black bars indicate the number of genes that are glucose- dependent and CovRS-dependent; grey bars indicate the number of genes that are glucose-dependent and CovRS-independent.</p

Additional file 1: of Negatively charged AuNP modified with monoclonal antibody against novel tumor antigen FAT1 for tumor targeting

Supplementary figures. (DOC 2242 kb

Graphical representation summarizing adaptive regulation of GBS in high glucose conditions.

<p>Genes of interest are color-grouped according to main functional categories. Arrows indicate up- or down-regulation relative to time of 30′ in high glucose <i>vs.</i> no glucose.</p

List of genes highly regulated in GBS strain 2603V/R after incubation in high glucose medium.

<p>List of genes highly regulated in GBS strain 2603V/R after incubation in high glucose medium.</p

Enhancing Antibody Serodiagnosis Using a Controlled Peptide Coimmobilization Strategy

Antigen immunoreactivity is often determined by surface regions defined by the 3D juxtapositions of amino acids stretches that are not continuous in the linear sequence. As such, mimicking an antigen immunoreactivity by means of putative linear peptide epitopes for diagnostic purposes is not trivial. Here we present a straightforward and robust method to extend the reach of immune-diagnostic probes design by copresenting peptides belonging to the same antigenic surface. In this case study focused on a computationally predicted Zika virus NS1 protein putative antigenic region, we reached a diagnostic confidence by the oriented and spatially controlled coimmobilization of peptide sequences found adjacent within the protein fold, that cooperatively interacted to provide enhanced immunoreactivity with respect to single linear epitopes. Through our method, we were able to differentiate Zika infected individuals from healthy controls. Remarkably, our strategy fits well with the requirements to build high-throughput screening platforms of linear and mixed peptide libraries, and it could possibly facilitate the rapid identification of conformational immunoreactive regions