Project description:Biomanufacturing remains financially uncompetitive with the lower cost but higher carbon emitting hydrocarbon based chemical industry. Novel chassis organisms may enable cost reductions with respect to traditional chassis such as E. coli and so open an economic rout to low emission biomanufacturing. Extremophile bacteria exemplify that potential. Salt tolerant halomonas species thrive in conditions inimical to other organisms. Their adoption would eliminate the cost of sterilising equipment. Novel chassis are inevitably poorly understood in comparison to established organisms. Rapid characterisation and community data sharing will facilitate organisms’ adoption for biomanufacturing. This paper describes baseline proteomics data set for Halomonas bluephagenesis TD01 under active development for biomanufactoring. The data record comprises a newly sequenced genome for the organism; evidence for expression of 1150 proteins (30% of the proteome) including baseline quantification of 1050 proteins (27% of the proteome) and a spectral library enabling re-use for targeted proteomics assays. Protein data is annotated with KEGG Orthology enabling rapid matching of quantitative data to pathways of interest to biomanufacturing.
Project description:PU.1 is an Ets family transcription factor that is essential for the differentiation of both myeloid and lymphoid cells. PU.1 is down-regulated in classical Hodgkin lymphoma cells via methylation of the PU.1 promoter. To evaluate whether down-regulation of PU.1 is essential for the growth of cHL cells, we generated KM-H2 derived cell lines conditionally express PU.1 by tet-off system (designated KM-H2tetPU.1). Conditonally expressed PU.1 by tetracycline removal induced complete growth arrest and apoptosis in KM-H2 cells. To elucidate the mechanisms underlying cell cycle arrest and apoptosis induced by PU.1, we compared gene expression profiles of KM-H2tetPU.1 cells 0, 1 and 3 days after PU.1 induction, by DNA microarray.
Project description:Rhodamin 123 is a dye which can be used to detect the activity of ABC transporters. We observed that after staining of KM-H2 Hodgkin lymphoma cells with Rhodamin 123, part of the cells rapidly eliminated the dye, while another part of the cells retained the dye for a longer time. We compared the transcriptome of KM-H2 Hodgkin lymphoma cells with high Rhodamin 123 efflux capacity and KM-H2 cells with low Rhodamin 123 efflux capacity.
Project description:PU.1 is an Ets family transcription factor that is essential for the differentiation of both myeloid and lymphoid cells. PU.1 is down-regulated in classical Hodgkin lymphoma cells via methylation of the PU.1 promoter. To evaluate whether down-regulation of PU.1 is essential for the growth of cHL cells, we generated KM-H2 derived cell lines conditionally express PU.1 by tet-off system (designated KM-H2tetPU.1). Conditonally expressed PU.1 by tetracycline removal induced complete growth arrest and apoptosis in KM-H2 cells. To elucidate the mechanisms underlying cell cycle arrest and apoptosis induced by PU.1, we compared gene expression profiles of KM-H2tetPU.1 cells 0, 1 and 3 days after PU.1 induction, by DNA microarray. We extracted total RNA from KM-H2tetPU.1 cells 0, 1 and 3 days after PU.1 induction by tetracycline removal. We compared gene expression profiles of KM-H2tetPU.1 cells 0, 1 and 3 days after PU.1 induction using DNA microarray analysis. 4 independent experiments were performed with each RNA samples.