Project description:A procyclic form Trypansome brucei RNAi line (PTT parental line, transfected with pALC14 incorporating a TbNMD3 gene fragment) capable of inducing depletion of TbNMD3 was analysed for mRNA expression by RNAseq Cells were grown for 72 hours in culture; RNAi was induced in cells by the addition of 1 microgram/ml of tetracycline
Project description:A procyclic form Trypansome brucei RNAi line (PTT parental line, transfected with pALC14 incorporating a TbNMD3 gene fragment) capable of inducing depletion of TbNMD3 was analysed for mRNA expression by RNAseq
Project description:RBSR2-TAP expressing cells were transfected with RNAi targeting RBSR2. 3 independent clones were used and RNAi was induced for 24 hours with 100ng/ml tetracycline. Cell lines expressing RBSR2-CTAP with no RNAi construct were used as controls. These are actually paired-end reads, file names ending _1 are one strand and _2 are the other strand.
Project description:RBSR1-CTAP expressing cells were transfected with RNAi targeting RBSR1. 3 independent clones were used and RNAi was induced for 24 hours with 100ng/ml tetracycline. PolyA enriched samples are paired-end reads, file names ending _1 are one strand and _2 are the other strand. rRNA depleted sample are single end reads.
Project description:How flagellar signaling regulates the host interaction of parasites remains a challenge due to poor conservation of signaling systems with those in cilia of higher organisms. The trypanosome-specific cAMP response protein 3 (CARP3) shows developmentally regulated localization at the flagellar tip membrane, where it is essential for parasite swarming and colonization of the tsetse fly insect vector. This project describes identification of CARP3-YFP interacting proteins by GFP trap pull down followed by mass spectrometry in the bloodstream stage as well as in the procyclic stage of Trypanosoma brucei.
Project description:Bloodstream-form trypanosomes with inducible RNAi targeting ZC3H28 were incubated with the tetracycline inducer for 10h, cells without tetracycline were used as controls. This experiment was done in triplicates. In another similar experiment, bloodstream-form trypanosomes with inducible RNAi targeting ZC3H28 were incubated with the tetracycline inducer for 14h, 16h, and 24h. Cells without tetracycline (named ''no tet'' or ''0h'') were used as controls. This experiment was done in duplicates.
Project description:Epitope tagging and magnetic bead enrichment were combined with SILAC quantitative proteomics to determine a high-confidence glycosome proteome for the procyclic life cycle stage of the parasite Trypanosoma brucei.