LD50 of <i>S. suis</i> type 2 HA9801strains and recombinant SS2 Egfp-HA9801 strains in Balb/C mice.

<p>The LD50 of both strains were calculated according to the Karber method.</p

Bacterial distribution in different organs from mice infected i.p. with HA9801 parent and Egfp-HA9801 recombinant strains.

<p>Bacterial loads in the blood (A) are expressed as CFU/ml, and in the liver (B), spleen(C), lung (D), kidney (E) and brain (F) as CFU/0.05 g of tissue. Results are expressed as mean ± SEM of at least three infected mice per p.i. time point. No significant differences were found between the two strains throughout the experiment (<i>P</i>>0.05).</p

Virulence of the <i>S. suis</i> parent strain HA9801 and Egfp-HA9801 recombinant in Balb/C mice.

<p>These measurements were performed over a period of 7-day post-infection.</p>a<p>Percentage of mice with clinical symptoms.</p>b<p>Percentage of mice that died due to infection.</p

HIV-1 Vif proteins with mutations that abolish the interaction with CBF-β and/or CUL5 are defective in degrading A3F.

<p>HEK293T cells were transfected with the control vector, WT Vif or a Vif mutant as indicated and harvested after 48-HA, tubulin and A3F-V5. Error bars indicate the standard deviation from triplicate experiments.</p

Epifluorescence microscopy analysis of bacteria cultured <i>in vitro</i>.

<p>A: Egfp-HA9801(10×40); B: HA9801(10×40); C: Egfp-HA9801(10×100); D: HA9801(10×100).</p

Epifluorescence microscopy analysis of the cryosections of the tissues obtained from the mice infected with Egfp-HA9801 or HA9801.

<p>The cryosections of liver (A-1), lung (B-1), kidney (C-1), spleen (D-1) and brain (E-1) from the Egfp-HA9801-infected mice were detected under epifluorescent microscope and the FITC images are shown in the left column. The FITC images of the cryosections of liver (A-2), lung (B-2), kidney (C-2), spleen (D-2) and brain (E-2) from the HA9801-infected mice, are showed in the right column.</p

Identification of two highly conserved regions in Vif proteins of various HIV-1 subtypes.

<p>(A) Conserved amino acids in Vif molecules. (B) Illustration of Vif mutant constructs.</p

Bacterial strains, and plasmids used in this study.

<p>Cm^r, chloromycetin resistant; Spc^r, spectinomycin resistant.</p

Genomic organization of the double crossover recombination locus in <i>S. suis</i> 2 Egfp-HA9801 and confirmation analysis of the recombinant strain Egfp-HA9801.

<p>(A) Genomic organization of the double crossover recombination locus and its flanking genes in <i>S. suis</i> 2 Egfp-HA9801. Dashes above the gene indicate restriction sites. The numbers above the gene and between the solid arrows indicate the size (bps) of the known gene fragments. The different color boxes represent <i>sly, egfp,spc^r</i> and <i>bsly</i> genes. The location of the primers used in PCR and RT-PCR detection are indicated by inverted arrowheads. (B) PCR analysis of the Egfp-HA9801 recombinant strain. The PCR primer combinations are shown over the lanes. Genomic DNA from the parent strain HA9801 (lane 2) and Egfp-HA9801 recombinant (lane 2) were used as templates. Lane 1 is the negative control. The 15 kb DNA ladder marker is shown to the left (M). (C) RT- PCR analysis of the Egfp-HA9801 recombinant strain. The primer combinations used in RT-PCR are shown over the lanes. Total RNA extracted from mid-exponential-phase cultures of the following strains were used as templates: parent strain HA9801 (lane 2, 5) and Egfp-HA9801 recombinant (lane 1, 4). Lane 3 and lane 6 are negative controls. The 15 kb DNA ladder marker is shown to the left (M). Theoretical size (bp) of each of the PCR and RT-PCR products generated with the primer combinations are shown in (A).</p

Sensitive Electrochemical Aptamer Biosensor for Dynamic Cell Surface <i>N</i>‑Glycan Evaluation Featuring Multivalent Recognition and Signal Amplification on a Dendrimer–Graphene Electrode Interface

We demonstrate a multivalent recognition and highly selective aptamer signal amplification strategy for electrochemical cytosensing and dynamic cell surface <i>N</i>-glycan expression evaluation by the combination of concanavalin A (Con A), a mannose binding protein, as a model, conjugated poly­(amidoamine) dendrimer on a chemically reduced graphene oxide (rGO–DEN) interface, and aptamer- and horseradish peroxidase-modified gold nanoparticles (HRP–aptamer–AuNPs) as nanoprobes. In this strategy, the rGO–DEN can not only enhance the electron transfer ability but also provide a multivalent recognition interface for the conjugation of Con A that avoids the weak carbohydrate–protein interaction and dramatically improves the cell capture efficiency and the sensitivity of the biosensor for cell surface glycan. The high-affinity aptamer- and HRP-modified gold nanoparticles provide an ultrasensitive electrochemical probe with excellent specificity. As proof-of-concept, the detection of CCRF-CEM cell (human acute lymphoblastic leukemia) and its surface <i>N</i>-glycan was developed. It has demonstrated that the as-designed biosensor can be used for highly sensitive and selective cell detection and dynamic evaluation of cell surface <i>N</i>-glycan expression. A detection limit as low as 10 cells mL^–1 was obtained with excellent selectivity. Moreover, this strategy was also successfully applied for <i>N</i>-glycan expression inhibitor screening. These results imply that this biosensor has potential in clinical diagnostic and drug screening applications and endows a feasibility tool for insight into the <i>N</i>-glycan function in biological processes and related diseases