Experiment Overview
| Repository ID: | FR-FCM-ZZQM | Experiment name: | Heterogeneity and plasmid copy number of E. coli carrying pSEVA plasmids | MIFlowCyt score: | 87.94% |
| Primary researcher: | Michael Jahn | PI/manager: | Michael Jahn | Uploaded by: | Michael Jahn |
| Experiment dates: | 2015-06-01 - 2015-12-31 | Dataset uploaded: | Apr 2016 | Last updated: | Nov 2016 |
| Keywords: | [reporter] [Fluorescent proteins] [EGFP] [plasmid] [plasmid copy number] [Heterologous expression] [Escherichia coli] [replication system] | Manuscripts: | |||
| Organizations: |
Helmholtz-Centre for Environmental Research - UFZ, Env. microbiology, Leipzig, (Germany)
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| Purpose: | The aim was to test performance and identify population heterogeneity in a set of standardized plasmids from the SEVA repository. The seven plasmids differed in their replication system, while the plasmid backbone containing lacIq/Ptrc and an EGFP reporter remained identical for all. | ||||
| Conclusion: | Six out of the seven constructs showed homogeneous distribution. Replication system 5 (pSEVA254-AEB, RSF1010 replication origin) showed a considerable amount of heterogeneity in terms of EGFP expression. Two exemplary populations were sorted (pSEVA244-AEB, pSEVA254-AEB) and the plasmid copy number determined on the sub-population level showed extreme variation. For details regarding study concept and sample description, see: Jahn et al., Microbial Cell Factories, 2016. | ||||
| Comments: | None | ||||
| Funding: | This work was supported by the European Union Regional Development Fund (ERDF) and the Free State of Saxony (Germany). | ||||
| Quality control: | Non-fluorescent control strains carrying EGFP-free plasmids were used as a control. | ||||
Experiment variables
| Timepoints | |
|---|---|
| · 0 | ATH_02_Nov_15_08.fcs · ATH_02_Nov_15_09.fcs · ATH_02_Nov_15_10.fcs · ATH_02_Nov_15_11.fcs · ATH_02_Nov_15_12.fcs · ATH_02_Nov_15_13.fcs · ATH_02_Nov_15_14.fcs · AMJ_23_Jun_15_13.fcs · AMJ_23_Jun_15_14.fcs · AMJ_23_Jun_15_16.fcs · AMJ_23_Jun_15_17.fcs · AMJ_23_Jun_15_18.fcs · AMJ_23_Jun_15_19.fcs · ATH_03_Dec_15_08.fcs |
| · 4 | ATH_02_Nov_15_15.fcs · ATH_02_Nov_15_16.fcs · ATH_02_Nov_15_17.fcs · ATH_02_Nov_15_18.fcs · ATH_02_Nov_15_19.fcs · ATH_02_Nov_15_20.fcs · ATH_02_Nov_15_21.fcs · AMJ_23_Jun_15_20.fcs · AMJ_23_Jun_15_21.fcs · AMJ_23_Jun_15_23.fcs · AMJ_23_Jun_15_24.fcs · AMJ_23_Jun_15_25.fcs · AMJ_23_Jun_15_26.fcs · ATH_03_Dec_15_12.fcs |
| · 8 | ATH_02_Nov_15_22.fcs · ATH_02_Nov_15_23.fcs · ATH_02_Nov_15_24.fcs · ATH_02_Nov_15_25.fcs · ATH_02_Nov_15_26.fcs · ATH_02_Nov_15_27.fcs · ATH_02_Nov_15_28.fcs · AMJ_23_Jun_15_27.fcs · AMJ_23_Jun_15_28.fcs · AMJ_23_Jun_15_30.fcs · AMJ_23_Jun_15_31.fcs · AMJ_23_Jun_15_32.fcs · AMJ_23_Jun_15_33.fcs · ATH_03_Dec_15_16.fcs |
| · 24 | ATH_02_Nov_15_29.fcs · ATH_02_Nov_15_30.fcs · ATH_02_Nov_15_31.fcs · ATH_02_Nov_15_32.fcs · ATH_02_Nov_15_33.fcs · ATH_02_Nov_15_34.fcs · ATH_02_Nov_15_35.fcs · AMJ_23_Jun_15_34.fcs · AMJ_23_Jun_15_35.fcs · AMJ_23_Jun_15_37.fcs · AMJ_23_Jun_15_38.fcs · AMJ_23_Jun_15_39.fcs · AMJ_23_Jun_15_40.fcs · ATH_03_Dec_15_20.fcs |
| Conditions | |
|---|---|
| · p224-AEB | ATH_02_Nov_15_08.fcs · ATH_02_Nov_15_15.fcs · ATH_02_Nov_15_22.fcs · ATH_02_Nov_15_29.fcs · AMJ_23_Jun_15_13.fcs · AMJ_23_Jun_15_20.fcs · AMJ_23_Jun_15_27.fcs · AMJ_23_Jun_15_34.fcs |
| · p234-AEB | ATH_02_Nov_15_09.fcs · ATH_02_Nov_15_16.fcs · ATH_02_Nov_15_23.fcs · ATH_02_Nov_15_30.fcs · AMJ_23_Jun_15_14.fcs · AMJ_23_Jun_15_21.fcs · AMJ_23_Jun_15_28.fcs · AMJ_23_Jun_15_35.fcs |
| · p244-AEB | ATH_02_Nov_15_10.fcs · ATH_02_Nov_15_17.fcs · ATH_02_Nov_15_24.fcs · ATH_02_Nov_15_31.fcs · ATH_03_Dec_15_08.fcs · ATH_03_Dec_15_12.fcs · ATH_03_Dec_15_16.fcs · ATH_03_Dec_15_20.fcs |
| · p254-AEB | ATH_02_Nov_15_11.fcs · ATH_02_Nov_15_18.fcs · ATH_02_Nov_15_25.fcs · ATH_02_Nov_15_32.fcs · AMJ_23_Jun_15_16.fcs · AMJ_23_Jun_15_23.fcs · AMJ_23_Jun_15_30.fcs · AMJ_23_Jun_15_37.fcs |
| · p264-AEB | ATH_02_Nov_15_12.fcs · ATH_02_Nov_15_19.fcs · ATH_02_Nov_15_26.fcs · ATH_02_Nov_15_33.fcs · AMJ_23_Jun_15_17.fcs · AMJ_23_Jun_15_24.fcs · AMJ_23_Jun_15_31.fcs · AMJ_23_Jun_15_38.fcs |
| · p274-AEB | ATH_02_Nov_15_13.fcs · ATH_02_Nov_15_20.fcs · ATH_02_Nov_15_27.fcs · ATH_02_Nov_15_34.fcs · AMJ_23_Jun_15_18.fcs · AMJ_23_Jun_15_25.fcs · AMJ_23_Jun_15_32.fcs · AMJ_23_Jun_15_39.fcs |
| · p284-AEB | ATH_02_Nov_15_14.fcs · ATH_02_Nov_15_21.fcs · ATH_02_Nov_15_28.fcs · ATH_02_Nov_15_35.fcs · AMJ_23_Jun_15_19.fcs · AMJ_23_Jun_15_26.fcs · AMJ_23_Jun_15_33.fcs · AMJ_23_Jun_15_40.fcs |
| Sample Type | |
|---|---|
| · R1 | ATH_02_Nov_15_08.fcs · ATH_02_Nov_15_09.fcs · ATH_02_Nov_15_10.fcs · ATH_02_Nov_15_11.fcs · ATH_02_Nov_15_12.fcs · ATH_02_Nov_15_13.fcs · ATH_02_Nov_15_14.fcs · ATH_02_Nov_15_15.fcs · ATH_02_Nov_15_16.fcs · ATH_02_Nov_15_17.fcs · ATH_02_Nov_15_18.fcs · ATH_02_Nov_15_19.fcs · ATH_02_Nov_15_20.fcs · ATH_02_Nov_15_21.fcs · ATH_02_Nov_15_22.fcs · ATH_02_Nov_15_23.fcs · ATH_02_Nov_15_24.fcs · ATH_02_Nov_15_25.fcs · ATH_02_Nov_15_26.fcs · ATH_02_Nov_15_27.fcs · ATH_02_Nov_15_28.fcs · ATH_02_Nov_15_29.fcs · ATH_02_Nov_15_30.fcs · ATH_02_Nov_15_31.fcs · ATH_02_Nov_15_32.fcs · ATH_02_Nov_15_33.fcs · ATH_02_Nov_15_34.fcs · ATH_02_Nov_15_35.fcs |
| · R2 | AMJ_23_Jun_15_13.fcs · AMJ_23_Jun_15_14.fcs · AMJ_23_Jun_15_16.fcs · AMJ_23_Jun_15_17.fcs · AMJ_23_Jun_15_18.fcs · AMJ_23_Jun_15_19.fcs · AMJ_23_Jun_15_20.fcs · AMJ_23_Jun_15_21.fcs · AMJ_23_Jun_15_23.fcs · AMJ_23_Jun_15_24.fcs · AMJ_23_Jun_15_25.fcs · AMJ_23_Jun_15_26.fcs · AMJ_23_Jun_15_27.fcs · AMJ_23_Jun_15_28.fcs · AMJ_23_Jun_15_30.fcs · AMJ_23_Jun_15_31.fcs · AMJ_23_Jun_15_32.fcs · AMJ_23_Jun_15_33.fcs · AMJ_23_Jun_15_34.fcs · AMJ_23_Jun_15_35.fcs · AMJ_23_Jun_15_37.fcs · AMJ_23_Jun_15_38.fcs · AMJ_23_Jun_15_39.fcs · AMJ_23_Jun_15_40.fcs · ATH_03_Dec_15_08.fcs · ATH_03_Dec_15_12.fcs · ATH_03_Dec_15_16.fcs · ATH_03_Dec_15_20.fcs |
| · controls | ATH_02_Nov_15_03.fcs · ATH_02_Nov_15_04.fcs · ATH_02_Nov_15_06.fcs · AMJ_23_Jun_15_03.fcs · AMJ_23_Jun_15_04.fcs · AMJ_23_Jun_15_06.fcs |
