Journal: Viruses

Article Title: Simultaneous Production of a Virus-Like Particle Linked to dsRNA to Enhance dsRNA Delivery for Yellow Head Virus Inhibition

doi: 10.3390/v14122594

Figure Lengend Snippet: Construction of recombinant pET28a-Linked cp Pst DNV-ds pro vector and expression of its counterpart. ( A ) Schematic illustration for recombinant pET28a-Linked cp Pst DNV-ds pro . Asterisk indicates a stop codon. ( B ) SDS-PAGE and western blot analysis of total cell lysate from induced recombinant E. coli strain Rosetta-gami 2(DE3) pLysS. Lane M, PageRuler™ Prestained Protein Ladder (Thermo Fisher Scientific, Waltham, MA, USA). ( C ) The dsRNA-YHV protease gene expression and its properties. Total RNA isolated from induced bacterial cells was treated with RNase III or RNase A and analyzed on 10% TAE buffered-polyacrylamide gel. An arrow indicates ds pro . Lane M, GeneRuler 100 bp DNA Ladder (Thermo Fisher Scientific, Waltham, MA, USA).

Article Snippet: The plasmid DNA pellet was dissolved in 1.2 M NaCl containing 10 µg/mL RNase A (New England Biolabs, Ipswich, MA, USA) followed by incubation at 37 °C for 30 min. An equal volume of chloroform was added and mixed and the solution was centrifuged at 18,890× g for 15 min at 4 °C.

Techniques: Recombinant, Plasmid Preparation, Expressing, SDS Page, Western Blot, Isolation

Journal: Viruses

Article Title: Simultaneous Production of a Virus-Like Particle Linked to dsRNA to Enhance dsRNA Delivery for Yellow Head Virus Inhibition

doi: 10.3390/v14122594

Figure Lengend Snippet: Characterization of recombinant Linked cp Pst DNV-ds pro counterpart. ( A ) Transmission electron micrograph of Linked cp Pst DNV-ds pro (Linked Pst DNV VLPs-ds pro ). ( B ) Linked Pst DNV VLPs-ds pro after 6-month storage at 4 °C. ( C ) The presence of dsRNA on the outer surface of Linked Pst DNV VLPs-ds pro . Total RNA isolated from purified recombinant VLP was directly treated with either RNase III, RNase A or DNase I. ( D ) The encapsulated dsRNA in Linked Pst DNV VLPs-ds pro . Intrinsic dsRNA was confirmed by RNA isolation of RNase III-treated Linked Pst DNV VLPs-ds pro , followed by incubation with either RNase III, RNase A, or DNase I. All RNA products were analyzed on a 12% TAE buffered-polyacrylamide gel. Lane M, GeneRuler 100 bp DNA Ladder (Thermo Fisher Scientific, Waltham, MA, USA).

Article Snippet: The plasmid DNA pellet was dissolved in 1.2 M NaCl containing 10 µg/mL RNase A (New England Biolabs, Ipswich, MA, USA) followed by incubation at 37 °C for 30 min. An equal volume of chloroform was added and mixed and the solution was centrifuged at 18,890× g for 15 min at 4 °C.

Techniques: Recombinant, Transmission Assay, Isolation, Purification, Incubation

Journal: PLOS Biology

Article Title: Disruption of tRNA biogenesis enhances proteostatic resilience, improves later-life health, and promotes longevity

doi: 10.1371/journal.pbio.3002853

Figure Lengend Snippet: Volcano plots of differential expression of short RNAs (<300 b) in the superior duodenum between Polr3B -/+ and wild-type mice, with different classes of transcripts coloured in different panels. ( A ) Female. ( B ) Male. “Other” refers to 7SK , RNase P , MRP , U6 , Vault , and Y RNA; horizontal line indicates the p value threshold for 10% FDR. ( C ) Thapsigargin LD50 of primary fibroblasts in culture, isolated from female or male, wild-type or Polr3B -/+ mice ( n = 5–7, effect of genotype p = 6 × 10 −3 , effect of sex or interaction p < 0.05, LM ). Boxplots show quantiles with individual data points overlayed. Data underlying this figure can be found in .

Article Snippet: Sample preparation and sequencing for Ribo-Seq and the associated RNA-Seq was performed by CD Genomics, using 300 to 500 mg of whole flies, as described [ ] and using RNase I. Library was prepared with NEBNext Multiple Small RNA Library Prep Set for Illumina (catalogue no. E7300S, E7300L) and sequenced using Illumina HiSeq TM X10.

Techniques: Expressing, Isolation