Part:BBa_K4817013:Design
MomL: AHL lactonase
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 804
Illegal PstI site found at 917 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 804
Illegal PstI site found at 917 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 804
Illegal PstI site found at 917 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 804
Illegal PstI site found at 917 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The coding sequences of MomL and Del-MomL were connected to LacO/LacI (BBa_K1624002, BBa_K3257045) and pT7 (BBa_K4609008). IPTG was used to induce protein expression, simulating quorum sensing-induced protein expression to verify the function efficiency of the MomL. LacO/LacI are commonly found in the pET plasmids. IPTG (isopropyl β-D-1-thiogalactopyranoside) is a molecular analogue of allolactose and has the same function as allolactose. Both can act as inducers and bind to the repressor in the Lac operon, thereby preventing LacI from binding to LacO upstream of pT7 and ultimately initiating the expression of MomL. MomL crude extract was collected and used to treat E.coli DH5α(with p15A-lux-sfGFP). Without MomL, the LuxR secreted by J23100 in plasmid p15A (BBa_C0062) interacts with AHLs and initiates LuxP, expressing the green fluorescent protein sfGFP to emit fluorescence. When MomL exists, MomL degrades AHLs, LuxP cannot turn on the expression of sfGFP, and the green fluorescence weakens.
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Source
Muricauda olearia Th120