Team:MichiganState/Contribution
Characterization / Contribution
Characterization of the burden of BBa_K819010 in E. coli JEB1203 Burden Monitor Strain
The characterization of the part BBa_K819010 was added here.
Using a strain that UT Austin iGEM team made, JEB1203, we can measure the burden of a plasmid. This works because JEB1203 has GFP in the chromosome and when a plasmid that has a high burden, or takes a lot of energy from chromosomal replication/transcription, the GFP will not fluoresce as much. However, since most parts added are not necessary to the bacteria will get rid of the part or cause mutation to lighten that burden. From this information, we can tell whether or not a part has a high burden on the bacteria or not. This is how we decided the burden of part BBa_K819010.
We used the JEB1203 Burden monitor strain that has GFP in the chromosome to show how much of a burden part has on a cell. The higher the burden of a part means that the GFP and OD will be lower because the part is taking up more energy away from chromosomal replication and expression. This graph shows the expected OD and GFP levels of the JEB1203 E. coli strain with no burden.
This graph shows the OD and GFP fluorescence of the JEB1206 burden monitor E. coli strain, which has a medium/high burden. To be noted in this graph is the lower ODs and GFP levels than that of the monitor strain JEB1203, which has no burden.
This graph is the BBa_K819010 part that codes for cheZ transformed into the JEB1203 burden monitor E. coli strain. From this graph, it can be seen that the OD and GFP levels are at very similar levels of the JEB1203 strain that did not have any burden. From this data, we characterized the burden of BBa_K819010 as little to none.