Southern blot and northern blot analysis of shrimp Ago1 isoforms.

<p>(<b>A</b>) Southern blot of shrimp genomic DNA with DIG-labeled Ago1-probe that could detect three Ago1 isoforms or Ago1-fragment 2-probe that was unique to Ago1A and Ago1B. (<b>B</b>) Northern blot of total RNAs extracted from shrimp gills. The probes used were shown on the top. The upper band likely consisted of co-migrated Ago1A and Ago1B transcripts, while the lower band potentially represented the Ago1C transcript.</p

Additional file 1 of Radiochemotherapy-induced DNA repair promotes the biogenesis of gastric cancer stem cells

Additional file 1. Fig. S1: Heat map of the differentially expressed DNA repair genes between GCNSCs and cisplatin-induced, doxorubicin-induced or X-ray-induced GCSCs

Characterization of a novel non-specific nuclease from thermophilic bacteriophage GBSV1-3

C to 80°C. To evaluate the thermostability, the residual enzymatic activities were monitored after incubation of the enzyme solutions (295 μg/ml) in the absence of herring dsDNA (100 μg/ml) at 40, 50, 60 or 70°C (pH 7.5) for 15, 40, 60, 90 or 130 min. The nuclease activity assay was conducted using 100 μg/ml of herring dsDNA as substrate. Each point represented the mean of triplicate assays and the error bars showed the standard deviations.<p><b>Copyright information:</b></p><p>Taken from "Characterization of a novel non-specific nuclease from thermophilic bacteriophage GBSV1"</p><p>http://www.biomedcentral.com/1472-6750/8/43</p><p>BMC Biotechnology 2008;8():43-43.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2390534.</p><p></p

Characterization of a novel non-specific nuclease from thermophilic bacteriophage GBSV1-2

Se I (B, C and D) was also included as positive control. The protein solutions were shown on the top and the nucleic acids were indicated on the right. 1 μg of nucleic acids were respectively incubated with 1.5 μg of the purified GBSV1-NSN protein in 20 μl of reaction buffer at 37°C for six hours.<p><b>Copyright information:</b></p><p>Taken from "Characterization of a novel non-specific nuclease from thermophilic bacteriophage GBSV1"</p><p>http://www.biomedcentral.com/1472-6750/8/43</p><p>BMC Biotechnology 2008;8():43-43.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2390534.</p><p></p

Expression profiles of Ago1 isoforms in shrimp.

<p>(<b>A</b>) Expression patterns of Ago1 isoforms in different tissues or organs of shrimp as revealed by quantitative real-time PCR. The shrimp β-actin was used as an internal standard. The relative expression levels of Ago1A, Ago1B, and Ago1C mRNAs were compared with that of Ago1A in lymphoid organ. Each column represented the mean of triplicate assays within 1% standard deviation. (<b>B</b>) The time-course of expression profiles of Ago1 isoforms in lymphoid organ of shrimp challenged with WSSV by quantitative real-time PCR. The relative expression levels of Ago1A, Ago1B, and Ago1C mRNAs at various times post-inoculation (0, 12, 24, 48, and 72 h) were compared with that of Ago1A at 0 h post-inoculation. The numbers indicated the time points post-inoculation with WSSV. Each column represented the mean of triplicate assays within 1% standard deviation. The statistically significant differences between treatments were represented with an asterisk (*P<0.05).</p

Characterization of a novel non-specific nuclease from thermophilic bacteriophage GBSV1-7

, induced; 3, recombinant bacterium (containing gene), non-induced; 4, recombinant bacterium (containing gene), induced; 5, purified GST-GBSV1-NSN fusion protein; 6, purified GBSV1-NSN.<p><b>Copyright information:</b></p><p>Taken from "Characterization of a novel non-specific nuclease from thermophilic bacteriophage GBSV1"</p><p>http://www.biomedcentral.com/1472-6750/8/43</p><p>BMC Biotechnology 2008;8():43-43.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2390534.</p><p></p

Roles of Ago1 isoforms in the shrimp immune response against WSSV infection.

<p>To characterize the roles of Ago1 isoforms in the antiviral immunity, shrimp were injected with WSSV and isoform-specific siRNAs. Shrimp were injected simultaneously with WSSV and the low or high concentration of Ago1A-siRNA (<b>A</b>), Ago1B-siRNA (<b>B</b>), Ago1A/B-siRNA (<b>C</b>) or Ago1C-siRNA (<b>D</b>), respectively. As control, WSSV+control siRNA and WSSV only were included in the injections. At 48 h post-inoculation, the shrimp from each treatment were subjected to quantitative real-time PCR to quantify the expressions of Ago1A, Ago1B, and Ago1C (left). The solutions used for injections were shown in the box. At the same time, the WSSV loads in shrimp were monitored by quantitative real-time PCR (right). The statistically significant differences between treatments were represented with asterisk (*P<0.05). Lane headings showed the solutions used for injections.</p

DataSheet_1_Hallmarks of crustacean immune hemocytes at single-cell resolution.docx

In invertebrates, hemocytes are the key factors in innate immunity. However, the types of invertebrate immune hemocytes are unclassified due to the limitation of morphological classification. To determine the immune hemocytes of crustaceans, the heterogeneity of hemocytes of shrimp Marsupenaeus japonicus and crayfish Procambarus clarkii, two representative crustacean species, were characterized in this study. The results of single-cell RNA sequencing indicated that shrimp and crayfish contained 11 and 12 types of hemocytes, respectively. Each of different types of hemocytes specifically expressed the potential marker genes. Based on the responses of shrimp and crayfish to the infection of white spot syndrome virus (WSSV) and the challenge of lipopolysaccharide (LPS), four types of immune hemocytes of crustaceans were classified, including semi-granular hemocytes involved in antimicrobial peptide production, granular hemocytes responsible for the production of antimicrobial peptides, hemocytes related to cell proliferation and hemocytes in immunity-activated state. Therefore, our study provided the first classification of crustacean hemocytes as well as of immune hemocytes of crustaceans at the single-cell resolution, which would be helpful to understand the innate immunity of invertebrates.</p

Characterization of a novel non-specific nuclease from thermophilic bacteriophage GBSV1-8

Se I (B, C and D) was also included as positive control. The protein solutions were shown on the top and the nucleic acids were indicated on the right. 1 μg of nucleic acids were respectively incubated with 1.5 μg of the purified GBSV1-NSN protein in 20 μl of reaction buffer at 37°C for six hours.<p><b>Copyright information:</b></p><p>Taken from "Characterization of a novel non-specific nuclease from thermophilic bacteriophage GBSV1"</p><p>http://www.biomedcentral.com/1472-6750/8/43</p><p>BMC Biotechnology 2008;8():43-43.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2390534.</p><p></p

Contribution of the Argonaute-1 Isoforms to Invertebrate Antiviral Defense

<div><p>Argonaute (Ago) protein, the central component of the RNA interference (RNAi) pathway, plays important roles in host innate antiviral immunity. Most organisms harbor a large number of different Ago proteins and isoforms; however, the roles of Ago isoforms in immune defense against pathogens remain unclear. In the present study, three Argonaute-1 (Ago1) isoforms, termed Ago1A, Ago1B, and Ago1C, were found in <em>Marsupenaeus japonicus</em> shrimp. Quantitative real-time PCR (polymerase chain reaction) revealed that isoforms Ago1A and Ago1B containing an insertion sequence in the PIWI domain, were significantly up-regulated in lymphoid organ and hemolymph, and also upon white spot syndrome virus (WSSV) challenge, indicating the involvement of Ago1A and Ago1B in antiviral immunity. The results showed that silencing of Ago1A with a sequence-specific siRNA led to a significant increase of WSSV loads. It was revealed that knockdown of Ago1B mRNA by 37–70% resulted in higher virus loads in shrimp. However, upon silencing Ago1B by more than 85%, a two-fold increase in Ago1A mRNA was observed but viral load was the same as untreated controls challenged with WSSV, suggesting that the simultaneous up-regulation of Ago1A might compensate for the loss of Ago1B. These data indicated that Ago1A played more important roles in the antiviral immune response than Ago1B. The simultaneous inhibition of Ago1A and Ago1B resulted in a greater increase in viral loads than Ago1A or Ago1B alone, indicating that Ago1A and Ago1B isoforms were involved in shrimp antiviral immunity. It was revealed that Ago1C had no effect on virus infection. Therefore, the current study presented the first report on the contribution of Ago isoforms in the invertebrate defense against virus infection.</p> </div