Part:BBa_K2150013
A fusion protein combining tetX(BBa_K2150101) with GFP(BBa_E0040)
This protein(refered to as tetX-GFP) combines a tetracycline-degrading enzyme (tetX monooxygenase, BBa_K2150101) and a green fluorescent protein (GFPmt3, BBa_E0040) with a 3*GGGGS linker. TetX-GFP has activities of both tetX and GFP. As a degradation enzyme, it can degrade tetracycline(tc) and its analogs (see BBa_K2150101), and can give the tetracycline resistance to E.coli. As a green fluorescent protein, it inherits the normal function of BBa_E0040. However, we noticed that when expressed under the same promoter, the fluorescence intensity(FI) of tetX-GFP was weaker than that of BBa_E0040 at the same growth stage of E.coli, and it took longer for E.coli expressing tetX-GFP to reach the same FI as E.coli expressing BBa_E0040. The reason for this may be that the expression level of tetX-GFP is lower than BBa_E0040 under the same promoter or that it takes longer for tetX-GFP to mature.
TetX-GFP can accurately reflect the expression level of tetracycline-degrading enzyeme and report whether our system is working (whether our bacteria is degrading tetracycline). When tetX-GFP is expressed under a tetracycline-regulated promoter such as BBa_R0040, it will become a visible tetracycline sensor in the presence of tetR (BBa_C0040), with no need for additional tetracycline-resistance genes.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 562
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1859
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