Characterization of encystation in Giardia intestinalis
Sammanfattning: Giardia intestinalis is a protozoan parasite causing the diarrheal disease called giardiasis that infects millions of people worldwide each year. The life cycle of Giardia intestinalis is characterized by two stages; the sturdy, infectious cyst and the vegetative, motile trophozoite. This project sought to investigate the regulation of the encystation process where trophozoites transform into cysts. Twelve genes with interesting transcriptomic profiles were chosen to study as putative transcription factors and regulators of encystation. These genes were cloned onto a plasmid with a Strep-Tag and transfected into Giardia intestinalis. The protein expression and localization was studied using immunofluorescence microscopy with antibodies against the Strep-Tag at different time points after inducing encystation. Although the project did not fully characterize these genes, protein expression was seen in all cases except two. Some proteins were seen localized to the nuclei and others had a localization pattern similar to the localization of cyst wall protein. In addition, a dramatic phenotype resembling cells going through programmed cell death was observed in one of the transfectants early in encystation and would be interesting to study further. The transfectant strains generated from this project remain interesting candidates to investigate as putative transcription factors.
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