Journal: PLoS Pathogens
Article Title: Multiple roles of PP2A binding motif in hepatitis B virus core linker and PP2A in regulating core phosphorylation state and viral replication
Figure Lengend Snippet: Effects of linker mutations on CCC DNA formation. HepG2 were co-transfected with indicated HBc expression constructs and the HBV genomic construct defective in HBc expression and HBV PF DNA was extracted from the transfected cells seven days after transfection. The extracted DNA was digested with Dpn I to degrade input plasmids ( A ), or Dpn I plus the exonuclease I and III to removal all DNA except closed circular DNA ( B ), before agarose gel electrophoresis and Southern blot analysis. Novel PF DNA smears detected from certain mutants are marked with the white asterisks to the left of the relevant lanes (S141D, S141R, L143A, TT146/147DD). PF-RC, PF-RC DNA; CCC, CCC DNA; cM, closed minus strand DNA. C. CCC DNA and PF-RC DNA signals of each mutant were compared with WT (top two panels). CCC DNA and PF-RC DNA are normalized to core RC DNA (middle two panels), and CCC DNA is normalized PF-RC DNA (bottom). All values from the WT were set to 1.0. *, P
Article Snippet: Where indicated, Dpn I (NEB) digestion, instead of TURBO DNase, was used to remove the transfected plasmid after DNA purification [ ].
Techniques: Countercurrent Chromatography, Transfection, Expressing, Construct, Agarose Gel Electrophoresis, Southern Blot, Mutagenesis