Journal: Journal of Virology
Article Title: Methylation Patterns of Papillomavirus DNA, Its Influence on E2 Function, and Implications in Viral Infection
Figure Lengend Snippet: Effect of the global CpG methylation on BPV1 E2-dependent transcriptional activation. (A) Schematic of BPV1 LCR, the BamC promoter, and luciferase reporter plasmid pKT260, illustrating the relative positions of the BPV1 sequences, the basic BamC promoter from EBV, and the firefly luciferase translational ORF. Asterisk, relative position of the E2BS. (B) Bar graph summarizing the luciferase activities measured in lysates from cell populations transfected with methylated or unmethylated pKT260. Eight micrograms of the plasmid pKT260 that had or had not been treated in vitro with CpG methylase was transfected into C33A cells by electroporation in the presence or absence of the BPV1 E2 expression vector pCGE2 (10 μg). Luciferase activities were measured 48 h posttransfection. All experiments were repeated at least three times. Bars (error bars, standard deviations): 1, no DNA transfection control; 2, methylated pKT260 alone; 3 methylated pKT260 plus pCGE2; 4, unmethylated pKT260 alone; 5, unmethylated pKT260 plus pCGE2.
Article Snippet: In vitro DNA methylation was accomplished with CpG methylase ( Sss I methyltransferase), by following the procedure recommended by New England Biolabs, the commercial provider of Sss I.
Techniques: CpG Methylation Assay, Activation Assay, Luciferase, Plasmid Preparation, Transfection, Methylation, In Vitro, Electroporation, Expressing