Determining The Extent Of Trypansoma Brucei Peroxin 13.2 (TbPEX13.2)’S Glucose-Dependent Localization Of The Glycosomes In This Organism

Annah Nieman

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Microbiology

Presentation Type

Mentored

Mentor

Mentor: Meredith Morris, Department of Genetics and Biochemistry, Clemson University

Abstract

African trypanosomiasis spreads throughout Sub-Saharan Africa with the aid of the tsetse fly. The untreated disease is fatal and the drugs that are currently in use are toxic. This research studies parasite-specific essential processes with the goal of identifying new drug targets. Glycosomes are single-membrane bounded organelles unique to the Trypanosoma parasites. Glycosomal composition varies in response to environmental conditions and the disruption of glycosomes is lethal to the parasite. Two proteins, Trypanosoma brucei Peroxin (TbPEX13.1) and TbPEX13.2 are involved in glycosomal protein import and the silencing of either gene is lethal. Previous research has shown that the protein TbPEX13.1 changes localization in response to changes in extracellular glucose, a nutrient that fluctuates throughout the parasite lifecycle. The overall purpose of this study was to determine the extent to which TbPEX13.2, like TbPEX13.1, exhibits glucose-dependent localization. To do so, the gene TbPEX13.2 was amplified via Polymerase Chain Reaction and ligated into the pGEM vector, transformed into Ecoli, and followed by the excision of the TbPEX13.2 gene. The DNA sequence of TbPEX13.2 was then successfully sequenced. Due to time constraints, it could not be ligated into the pXS2 expression vector. Therefore, the expression construct was not transfected into trypanosomes and stable cell lines were not selected and TbPEX13.2eYFP expression and localization was not determined. The cloning of TbPEX13.2 into the pGEM vector was successful and TbPEX13.2 was successfully excised back out of the pGEM vector. But due to time constraints it could not be ligated into the pXS2 expression vector.

Start Date

4-11-2015 9:00 AM

End Date

4-11-2015 9:15 AM

COinS
 
Apr 11th, 9:00 AM Apr 11th, 9:15 AM

Determining The Extent Of Trypansoma Brucei Peroxin 13.2 (TbPEX13.2)’S Glucose-Dependent Localization Of The Glycosomes In This Organism

African trypanosomiasis spreads throughout Sub-Saharan Africa with the aid of the tsetse fly. The untreated disease is fatal and the drugs that are currently in use are toxic. This research studies parasite-specific essential processes with the goal of identifying new drug targets. Glycosomes are single-membrane bounded organelles unique to the Trypanosoma parasites. Glycosomal composition varies in response to environmental conditions and the disruption of glycosomes is lethal to the parasite. Two proteins, Trypanosoma brucei Peroxin (TbPEX13.1) and TbPEX13.2 are involved in glycosomal protein import and the silencing of either gene is lethal. Previous research has shown that the protein TbPEX13.1 changes localization in response to changes in extracellular glucose, a nutrient that fluctuates throughout the parasite lifecycle. The overall purpose of this study was to determine the extent to which TbPEX13.2, like TbPEX13.1, exhibits glucose-dependent localization. To do so, the gene TbPEX13.2 was amplified via Polymerase Chain Reaction and ligated into the pGEM vector, transformed into Ecoli, and followed by the excision of the TbPEX13.2 gene. The DNA sequence of TbPEX13.2 was then successfully sequenced. Due to time constraints, it could not be ligated into the pXS2 expression vector. Therefore, the expression construct was not transfected into trypanosomes and stable cell lines were not selected and TbPEX13.2eYFP expression and localization was not determined. The cloning of TbPEX13.2 into the pGEM vector was successful and TbPEX13.2 was successfully excised back out of the pGEM vector. But due to time constraints it could not be ligated into the pXS2 expression vector.