TaqMan Probe Design for Analysis of Stage-specific Genes in Toxoplasma gondii
Advisor Information
Paul Davis
Location
Milo Bail Student Center Ballroom
Presentation Type
Poster
Start Date
8-3-2013 1:00 PM
End Date
8-3-2013 4:00 PM
Abstract
Toxoplasma gondii is a spore and cyst forming parasite, granting this species the ability to infect various organisms throughout its life cycle. Transmission can occur in individuals by way of inadvertent ingestion simply by cleaning a cat’s litter box, as well as infecting soil and water supplies by improper disposal of feline fecal matter. Infection in humans can also occur by way of ingesting undercooked meat, (namely pork, but others have been reported as well) contaminated by T. gondii bradyzoites, cysts that are formed in muscular tissue by the parasite. This is particularly problematic in pregnant women contracting T. gondii as it is readily diffused across the placenta thereby infecting the unborn fetus leading to fetal abnormalities and in some cases even death. Although there have been several advances in the fabrication of various antibiotics used to treat the tachyzoite stage in T. gondii, only the human immune system has been able to keep the bradyzoite stage in check. In immunosuppressed individuals however, this immunoresponse is less than ample; for example, Toxoplasmosis is one of the leading killers in AIDS patients. The Davis labs’ primary target is the bradyzoite stage of the T. gondii life cycle. Previous experiments have focused on building an index of genes in T. gondii pointedly focused on the comparison of those expressed solely during the tachyzoite phase, and less on the bradyzoite stage. TaqMan probes were developed to aid in this process by targeting the housekeeping gene (all stages), ribosomal protein L33, as well as the bradyzoite stage specific marker lactate dehydrogenase. In this way we are now able to quantitatively monitor the percentage of bradyzoites to tachyzoites in Toxoplasmosis cultures by the level of expression shown in each of the two aforementioned genes.
TaqMan Probe Design for Analysis of Stage-specific Genes in Toxoplasma gondii
Milo Bail Student Center Ballroom
Toxoplasma gondii is a spore and cyst forming parasite, granting this species the ability to infect various organisms throughout its life cycle. Transmission can occur in individuals by way of inadvertent ingestion simply by cleaning a cat’s litter box, as well as infecting soil and water supplies by improper disposal of feline fecal matter. Infection in humans can also occur by way of ingesting undercooked meat, (namely pork, but others have been reported as well) contaminated by T. gondii bradyzoites, cysts that are formed in muscular tissue by the parasite. This is particularly problematic in pregnant women contracting T. gondii as it is readily diffused across the placenta thereby infecting the unborn fetus leading to fetal abnormalities and in some cases even death. Although there have been several advances in the fabrication of various antibiotics used to treat the tachyzoite stage in T. gondii, only the human immune system has been able to keep the bradyzoite stage in check. In immunosuppressed individuals however, this immunoresponse is less than ample; for example, Toxoplasmosis is one of the leading killers in AIDS patients. The Davis labs’ primary target is the bradyzoite stage of the T. gondii life cycle. Previous experiments have focused on building an index of genes in T. gondii pointedly focused on the comparison of those expressed solely during the tachyzoite phase, and less on the bradyzoite stage. TaqMan probes were developed to aid in this process by targeting the housekeeping gene (all stages), ribosomal protein L33, as well as the bradyzoite stage specific marker lactate dehydrogenase. In this way we are now able to quantitatively monitor the percentage of bradyzoites to tachyzoites in Toxoplasmosis cultures by the level of expression shown in each of the two aforementioned genes.