tetC Genes Research Articles

Regulation of the “ tetCD” genes of transposon Tn 10

In addition to the genes involved in tetracycline resistance, the loop region of the composite transposon Tn 10 contains two other known genes, tetC and tetD, whose functions are unclear. Using primarily a genetic approach, we examined tetCD gene expression and regulation. The tetC gene product, TetC, is a diffusible repressor of both tetC and tetD transcription. Despite an earlier claim by others, we do not detect induction of either tetC or tetD by tetracycline (Tc) or several of its analogs. Although the 5′ ends of the tetC and tetD messages overlap due to transcription from convergent promoters, we find no evidence for anti-sense RNA control. The operator for the TetC repressor has been localized. We also demonstrate that transcription from the tetD promoter probably terminates within IS 10-Right and does not apparently interfere with Tn 10 or IS 10-Right transposition or its regulation.

Immunization of mice with DNA encoding fragment C of tetanus toxin

Immunization of mice with Fragment C protein, the non-toxic C-terminal domain of tetanus toxin, will protect mice against lethal challenge with tetanus toxin. A plasmid, pcDNA3/tetC, which encodes a synthetic tetC gene expressed under the control of the human cytomegalovirus major intermediate early promoterlenhancer region, was constructed. Fragment C expression was observed in Chinese hamster ovary cells following transfection with pcDNA3/tetC. The immune response induced by intramuscular immunization with pure pcDNA3/tetC DNA was evaluated in a murine model. Anti-Fragment C serum immunoglobulin and proliferative responses in splenocytes were observed following two immunizations with pcDNA3/tetC. The major IgG subclass that recognized Fragment C was IgG2a and the stimulated splenocytes secreted high levels of interferon-γ. Sufficient anti-Fragment C serum immunoglobulins were induced by DNA-mediated immunization to protect mice against lethal challenge with tetanus toxin.

Immune response in mice following immunization with DNA encoding fragment C of tetanus toxin.

Tetanus toxin is a potent neurotoxin synthesized by Clostridium tetani. Immunization with fragment C protein, the nontoxic C-terminal domain of tetanus toxin, will protect mice against lethal challenge with tetanus toxin. A synthetic gene encoding fragment C (tetC) had previously been shown to express high levels of fragment C in Saccharomyces cerevisiae. A plasmid, pcDNA3/tetC, which encodes the synthetic tetC gene expressed under the control of the human cytomegalovirus major intermediate-early promoter/enhancer region, was constructed. Expression of fragment C was observed in eukaryotic cells growing in vitro following transfection with pcDNA3/tetC. The immune response induced by intramuscular immunization with pure pcDNA3/tetC DNA was evaluated in a murine model. Anti-fragment C serum immunoglobulin and proliferative responses in splenocytes were observed in BALB/c mice following two immunizations with pcDNA3/tetC. The major immunoglobulin G subclass that recognized fragment C was immunoglobulin G2a, and the stimulated splenocytes secreted high levels of gamma interferon. Immunity to tetanus is dependent on the presence of neutralizing serum antibodies against tetanus toxin. Sufficient anti-fragment C serum immunoglobulins were induced by DNA-mediated immunization to protect mice against lethal challenge with tetanus toxin.