Article Title: Promoter Sequence of Shiga Toxin 2 (Stx2) Is Recognized In Vivo, Leading to Production of Biologically Active Stx2
Figure Lengend Snippet: (A) Mice displayed a lethal dose-related response to pStx2. Survival rates of mice injected with different doses of pStx2 are shown. Mice injected with pStx2ΔAB or pGEMT were used as negative controls. Mice injected with 1 LD 100 and 3 LD 100 of purified Stx2 were used as positive controls. (B) Stx2 mRNA was detected in the liver. A real-time-PCR curve using cDNA from livers of mice inoculated with pStx2 (dotted line) or pGEMT (gray line) is shown. As a positive control, 50 pg of plasmid pStx2 (bold line) was used. (C) DNA electrophoresis. Lane 1, 50-bp ladder; lane 2. cDNA from livers of mice inoculated with pStx2; lane 3, cDNA from livers of mice inoculated with pGEMT; lane 4, pStx2 was used as positive control.
Article Snippet: The pGEMT Easy vector carrying the sequence of Stx2 was digested and religated in order to delete the active site of Stx2, generating the plasmid pStx2ΔAB ( ) as a Stx2-specific biological activity control.
Techniques: Mouse Assay, Injection, Purification, Real-time Polymerase Chain Reaction, Positive Control, Plasmid Preparation, Nucleic Acid Electrophoresis