Part:BBa_K1675010
FimE recombinase device
FimE is different from other kinds of recombinase. Its recognized sites on the upstream (IRL) and downstream (IRR) are not same. It means that after the recombination mediated by FimE happens, the sign sites would be exchanged, so that the new sign sites couldn’t be recognized by FimE again. So the recombination mediated by FimE is irreversible. Granted that the bacteria produce the strong alkali in the primary, the environment would turn alkaline quickly. So P-atp2 can correspond the change of environment and start the expression of FimE. FimE recognizes IRL and IRR, then catalyze the inversion of J23119. The weak acid would be generated at that time. For the environment, due to the production of the weak acid, it would turn neutral gradually.
We constructed our verification circuits through OE-PCR, and linked them into vector pSB1C3 (Fig.1,2). And we divided it into recombinase part and fluorescence part.
Fig.1 The constructed result of FimE, P-atp2 and P-atp2+B0034+FimE(channel 1-3 are P-atp2, Bxb1 and P-atp2+B0034+Bxb1 respectively)
Fig.2 The construction result of FimE’s verification device K137058(channel 1~6 the construction result of K137058, channel 1-3 and 6 are positive.)
Meanwhile, we constructed the verification devices, fluorescence part, respectively, then co-transformed them into BMTOP10. We observed their fluorescent condition by fluorescence microscope and floe cytometry.
Fig.3 The fluorescence result of FimE’s verification device
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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