Coding

Part:BBa_K2279000

Designed by: Yizhen Xu   Group: iGEM17_TMMU-China   (2017-10-15)
Revision as of 01:44, 2 November 2017 by Huqiwen (Talk | contribs)


AimR

AimR is a transcription factor.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 429
    Illegal SapI.rc site found at 169
    Illegal SapI.rc site found at 492

Plasmid construction

We used PCR to produce AimR gene fragment.

T--TMMU-China--aimr1.jpeg

Then we inserted this gene to plasmid pSB1C3. We transformed this recombinant plasmid (one contains gene AimR) into strain DH5α (E. coli). Then we picked some colonies for cultivation and extracted the recombinant plasmid, which was verified by enzyme digestion using PstI and EcoRI enzymes. From the result of electrophoresis, we confirmed the construction of pSB1C3-AimR was succeeded.

T--TMMU-China--aim2.jpeg

We then sequenced the positive bacteria and confirmed that the plasmid plays (one contains gene AimR) was indeed transferred to the bacteria. We expanded the bacteria and extracted plasmids from the bacteria. So far, we've successfully constructed AimR.

Biological function

The B.subtilis bacteriophage phi3T encode the Aim system to make lysis-lysogeny decision. The AimR is a transcription factor, the AimP is the propeptide of the mature signal peptide. However, binding of AimP to the AimR will disrupt the dimer forms of AimR. After that, the AimR can no longer bind to the promoter of AimX, a potential non coding RNA involved in the process of lysis-lysogeny. T--TMMU-China--aimfu.jpeg

The AimR-AimP system and its role in the phage lysis-lysogeny decision, cited from reference [1].

Design

By combining the expression of AimR and AimP components, we want to develop a synthetic QS system in B.subtilis for target gene autoinhibition. AimAuto.jpeg

A synthetic communication pathway between B.subtilis strains by co-culturing AimP-producing “sender” cells with AimR-sensing “receiver” cells to inhibit gene expression was also designed.

AimSR.jpeg


Characterization

Aim-fluorescence副本.jpg

Reference

[1] Erez, Z., Steinberger-Levy, I., Shamir, M., Doron, S., Stokar-Avihail, A., Peleg, Y., Melamed, S., Leavitt, A., Savidor, A., Albeck, S., et al. (2017). Communication between viruses guides lysis-lysogeny decisions. Nature 541, 488-493.

[edit]
Categories
Parameters
None