Protein Interaction Between Basal Body and Kinetoplast DNA Complex Within Trypanosoma brucei

Trypanosoma brucei is responsible for the disease Human African Trypanosomiasis (HAT), also known as African Sleeping Sickness, in humans as well as N’gana in livestock. Our work was done on the sub-species T. brucei brucei, which is non-pathogenic in humans, and thus safe for undergraduate laboratory research. Trypanosoma brucei is part of a diverse group of flagellated protozoans known as the kinetoplastids. Kinetoplastids are named for the concentration of mitochondrial DNA located at the posterior end of the mitochondrion and adjacent to the flagellum, called the kinetoplast DNA (kDNA). The kinetoplast is connected to the basal body (motor) of the flagellum by proteins through both membranes of the mitochondrion. The flagella drive the process of binary fission (cell division), suggesting that the replicated kDNA is separated through these protein interactions. Specific proteins were tested for their participation in the connection between kDNA and basal body. Our genes of interest were inserted into plasmid vectors. The plasmids were then linearized, followed by transfection into trypanosomes. Immunofluorescence was used to identify where these proteins localized within the cell.