Part:BBa_K4645016
J23100-B0030-vqmA-B0017-Pqtip-tetR-B0015-PTet-amcyan-B0017
Verification of Not-Gate.
Usage and Biology
We transformed this biobrick into E. coli to regulate expression of genes. With the growing of bacteria, DPO accumulates. While the concentration of DPO reached the threshold value, the expression of NeuA, NeuB and TetR would be activated. Then TetR shut down the expression of AmCyan in downstream.
Characterization
To test whether this biobrick can work as expected, we built a circuit that vqmA is under the control of lac operator, then transformed into BL21. We cultured blank BL21 (which had never been transformed any plasmid) and BL21 contain this plasmid till the OD600 of bacteria medium reached 0.6. Then BL21 with the plasmid were induced with 0mM IPTG、1mM IPTG. Whereafter, these 3 groups of bacteria were cultured in microplate reader 37°C, 220 rpm for 3.5 hours and detected OD600, fluorescence intensity (458 nm excitation light, 489 nm emission light) every 10 minutes.
Unluckily, induced bacteria showed stronger fluorescence intensity than control one, which is contrary to the expected result. We speculate that it might lead by the malfunction of qtip promoter.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 599
Illegal AgeI site found at 2547 - 1000COMPATIBLE WITH RFC[1000]
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