Part:BBa_K1442100:Experience

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Applications of BBa_K1442100

Figure 6. Characterization of mRdRp and RdRp activity with S3, R5 and C2/HP 3’ UTRs. Data was collected at different time points using Tecan Microplate reader. Controls also present: (IPTG + RBS + GFP) and NEB only.

Table 1. RNA dependent RNA polymerase activity following induction at 0.2 and 0.4 OD, relative to control wells (not induced at 0.2 and 0.4 OD). Error bars indicate standard error.

Horizontal axis depicts various different points (1-9: due to the nature of the analysis time in seconds is not displayed). RdRp induction at 0.2 and 0.4 OD at points 1-3 indicates initial activity and relative increase in fluorescence output. Initial RdRp activity then decays following induction, with decrease in fluorescence activity at both OD 0.2 and 0.4. Conclusively, results suggest initial induction of RdRp activity causes a relative increase in fluorescence with a stabilization/decrease thereafter.

Figure 7. RNA dependent RNA polymerase activity following induction at 0.2 and 0.4 OD, relative to control wells (not induced at 0.2 and 0.4 OD).

Figure 8. RNA dependent RNA polymerase activity following induction at 0.2 and 0.4 OD. Rates were extracted by linear regression and is displayed. Error bars indicate standard error of the mean. Rdrp Ni: Non induced, Rdrp I: Induced, mRdRp follows the same conventions.

This part has been extensively tested by the Warwick iGEM Team 2014, we have managed to document this part extensively with the results obtained depicted above. In terms of optimizing the chances of obtaining stable transformants, we recommend following the protocols that are found on our website.

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