Difference between revisions of "Part:BBa K2279001"
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[[file:T--TMMU-China--aimP.jpeg]]
 
[[file:T--TMMU-China--aimP.jpeg]]
  
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Then we inserted this gene to plasmid pSB1C3. We transformed this recombinant plasmid (one contains gene AimP) into strain DH5α (<i>E. coli</i>). Then we picked some colonies for cultivation and extracted the recombinant plasmid, which was verified by PCR. From the result of electrophoresis, we confirmed the transformation of AimP was success.  
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Then we inserted this gene to plasmid pSB1C3. We transformed this recombinant plasmid (one contains gene AimP) into strain DH5α (<i>E. coli</i>). Then we picked some colonies for cultivation and extracted the recombinant plasmid, which was verified by PCR and sequencing. From the result of electrophoresis, we confirmed the transformation of AimP was success.  
  
 
[[file:T--TMMU-China--aimp4.jpeg]]
 
[[file:T--TMMU-China--aimp4.jpeg]]
  
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We then sequenced the positive bacteria and confirmed that the plasmid plays (one contains gene AimP) was indeed transferred to the bacteria. We expanded the bacteria and extracted plasmids from the bacteria. So far, we've successfully constructed AimP.
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===Biological Function===
 
===Biological Function===

Revision as of 02:17, 2 November 2017


AimP

AimP encodes a signal peptide.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 58
  • 1000
    COMPATIBLE WITH RFC[1000]

Plasmid construction

We used PCR to produce AimP gene fragment.

T--TMMU-China--aimP.jpeg

Then we inserted this gene to plasmid pSB1C3. We transformed this recombinant plasmid (one contains gene AimP) into strain DH5α (E. coli). Then we picked some colonies for cultivation and extracted the recombinant plasmid, which was verified by PCR and sequencing. From the result of electrophoresis, we confirmed the transformation of AimP was success.

T--TMMU-China--aimp4.jpeg


Biological Function

The B.subtilis bacteriophage phi3T employs the AimR-AimP QS system to make the lysis-lysogeny decision. AimP is the pre-pro-peptide. It is 43aa long. The mature signal peptide of AimP is SAIRGA. Binding of the mature signal peptide to 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

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

Figure 1. The design of autoinhibiton system

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

Figure 2. The design of AimR-AimP based Sender and Receiver cells.

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.