Pseudomonas exotoxin antisense RNA selectively kills hepatitis B virus infected cells

Figure 1 A: p92 has the promoter and stem loop sequence of the T7 phage gene 10 and the Fd phage transcription terminator. In p95, p100 and p77, the LacZ-gene NotI-fragment of p92 is replaced by fragments containing truncated mutated forms of Pseudomonas exotoxin (PE38). p92 represents the control β-Gal expressing plasmid. p95 contains the gene for PE38 in reverse orientation to the CMV promoter. p77 expresses a non-functional frameshift mutant of PE38. p100 expresses the fully active PE38. B: Reverse transcription of toxin antisense RNA in vitro. Ethidium bromide stained nondenaturing 2% agarose gel. PCR fragments indicate the presence of toxin cDNA; templates were p100 RNA incubated with (1) no M-MLV RT, no random hexamer primers, (2) no M-MLV RT, random hexamer primers, (3) M-MLV RT, no random hexamer primers, (4) M-MLV RT, random hexamer primers.