MOESM1 of A novel strategy to improve protein secretion via overexpression of the SppA signal peptide peptidase in Bacillus licheniformis

Additional file 1. All the sequences of the primers and concentrations of total intracellular proteins were listed in the Additional file (Table S1, Table S2 and Table S3). The agarose gel electrophoresis analysis of the SPPs deficient strains and the fluorescence detection of the cell and the fermentation supernatant of the BL10/pHY-GFP, BL10S/pHY-GFP, BL10GS/pHY-GFP were also contained in the Additional file (Fig. S1, Fig. S2 and Fig. S3). This information is available free of charge via the Internet http://microbialcellfactories.biomedcentral.com/

Cd treatment triggered liver apoptosis in purse red common carp.

<p>(A) White arrows indicate the brown TUNEL-positive cells (hepatocytes). Representative photographs are shown at 200 × magnification. (B) Quantitative analysis of the percentage of apoptotic cells. Data shown represent mean ± SD from 4 fish in each group, error bars indicate standard deviation. Statistical significance was analyzed using a factorial ANOVA (* <i>P</i> < 0. 05).</p

Primers used for caspase-3A gene cloning and expression analysis.

<p>Primers used for caspase-3A gene cloning and expression analysis.</p

Multiple alignment of caspase-3 amino acid sequences.

<p>Identical amino acids are indicated by asterisks, whereas those with high or low similarities are indicated by semicolons and dots. The putative cleavage sites at aspartic acid residues (D), which are shaded in gray, separate the pro-domain (<b><—></b>), the large subunit (<b><·····></b>) and the small subunit (<b><-----></b>) respectively. The caspase family signature and the pentapeptide active-site motif (QACRG) were boxed in a continuous and discontinuous line, respectively. The protein binding domain (GSWFI) and integrin recognition motif (RGD) are indicated by solid circles. The GenBank accession numbers of caspase-3 amino acid sequences used here are as follows: zebrafish caspase-3A NP_571952.1, zebrafish caspase-3B NP_001041531.1, Medaka caspase-3A NP_001098140.1, Medaka caspase-3B NP_001098168.1, human NP_116786.1, mouse NP_056548.2, chicken (<i>Gallus gallus</i>) NP_990056.1 and frog (<i>Xenopus laevis</i>) NP_001081225.1.</p

Immunolocalization of caspase-3A in liver sections.

<p>NC represents negative control (substitution normal non-immune serum from the same host animal as the primary antibody). All the negative controls show no positive staining [Fig. 7(A-C), 7(G-I)]. Both control groups [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083423#pone-0083423-g007" target="_blank">Fig 7(D and J)</a>] and experimental groups [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083423#pone-0083423-g007" target="_blank">Fig 7(E, F, K and L)</a>] reveal weak immunolabeling. Arrows indicate the cytoplasm of hepatocytes. The representative photographs are shown at 200 × magnification.</p

Effects of Cd treatment on caspase-3A activity in liver from purse red common carp.

<p>The extracts were incubated with the specific substrates of caspase-3 and release of <i>p</i>-nitroanilide was measured at 405 nm. Values represent the mean ± SD; error bars indicate standard deviation. Statistical significance was analyzed using a factorial ANOVA (* <i>P</i> < 0. 05).</p

Dissolvability analyses of fusion protein and protein expression analysis

<p>(A): common carp caspase-3A clones expressed in <i>E. coli.</i> Lane M: protein molecular standard; Lane 1: total cellular proteins of PGEX-GST/BL21 after induction; Lane 2: supernate of purified recombinant PGEX-CSP3A; Lane 3: precipitate of purified recombinant PGEX-CSP3A. (B): Western blots showing procaspase-3A/ (Pro-csp3) and cleaved fragments (p17 and p12) after Cd treatment. (C): Protein band density was analyzed with the Gel-Pro Analyzer. GAPDH was used as a loading control. Each value was expressed as the ratio of caspase-3A proenzyme and its activated form (the sum of expression levels of p17 and p12 forms) to GAPDH level, which represents the mean ± SD. of 4 independent samples performed in triplicate. Statistical analysis was performed using a factorial ANOVA. Different upper case letters represent significant differences of proenzyme levels between exposure concentrations (<i>P</i> < 0.05), while different lower case letters represent significant differences of activated forms between exposure concentrations (<i>P</i> < 0.05) and asterisk (*) shows the significant difference between the two exposure time points for the same Cd-induced concentration (<i>P</i> < 0.05)</p

Effect of Cd treatment on mRNA transcript level of caspase-3A.

<p>The changes of mRNA levels were determined by real-time quantitative PCR. β-actin was used as a reference gene to normalize caspase-3A. The relative expression ratio was calculated with the 2^−ΔΔCT method. Error bars indicate standard deviation. Values represent the mean ± SD. of 4 independent samples. Statistical significance (<i>P</i> < 0.05) was analyzed using a factorial ANOVA. The mRNA levels of caspase-3A in carp were not significantly increased after exposure to Cd (<i>P</i> > 0.05).</p

High-Performance, Solution-Processed Quantum Dot Light-Emitting Field-Effect Transistors with a Scandium-Incorporated Indium Oxide Semiconductor

Light-emitting field-effect transistors (LEFETs) have attained great attention due to their special characteristics of both the switching capacity and the electroluminescence capacity. However, high-performance LEFETs with high mobility, high brightness, and high efficiency have not been realized due to the difficulty in developing high electron and hole mobility materials with suitable band structures. In this paper, quantum dot hybrid LEFETs (QD-HLEFETs) combining high-luminous-efficiency quantum dots (QDs) and a solution-processed scandium-incorporated indium oxide (Sc:In₂O₃) semiconductor were demonstrated. The red QD-HLEFET showed high electrical and optical performance with an electron mobility of 0.8 cm² V^–1 s^–1, a maximum brightness of 13 400 cd/m², and a maximum external quantum efficiency of 8.7%. The high performance of the QD-HLEFET is attributed to the good energy band matching between Sc:In₂O₃ and QDs and the balanced hole and electron injection (less exciton nonradiative recombination). In addition, incorporation of Sc into In₂O₃ can suppress the oxygen vacancy and free carrier generation and brings about excellent current and optical modulation (the on/off current ratio is 10⁵ and the on/off brightness ratio is 10⁶)