dttp thermo fisher  (New England Biolabs)

Journal: Viruses

Article Title: Factor-Dependent Internal Ribosome Entry Site and -1 Programmed Frameshifting Signal in the Bemisia-Associated Dicistrovirus 2

doi: 10.3390/v16050695

Figure Lengend Snippet: BaDV-2 IGR supports internal ribosome entry. ( A ) Bicistronic reporter RNAs containing either the CrPV IGR or the BaDV-2 IGR, ( B ) bicistronic RNAs with a 5′UTR strong hairpin, or ( C ) monocistronic reporter RNAs with a 5′UTR hairpin and the BaDV-2 IGR (top schematics) were incubated in Sf-21 extracts. In vitro transcribed RNAs were analyzed by agarose gel analysis (middle). Translation of FLuc and RLuc was measured by either radioisotope [ 35 S]-methionine/cysteine incorporation followed by SDS-PAGE and phosphorimager analysis or luciferase assays (graphs shown bottom). A paired t -test was used to determine the p values. * * p < 0.01. “n.s.” denotes the difference is not significant between the control groups and the experimental groups ( p > 0.05). Shown are representative SDS-PAGE gels and the averages from at least three independent experiments ± standard deviation.

Article Snippet: Monocistronic, bicistronic, and infectious clones were linearized by NarI, BamHI, and Eco53KI (NEB) restriction enzymes, respectively.

Techniques: Incubation, In Vitro, Agarose Gel Electrophoresis, SDS Page, Luciferase, Control, Standard Deviation