Difference between revisions of "Part:BBa K2279001"

Revision as of 05:15, 1 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.

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

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.

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.

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.

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.

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.

@@ Line 17: / Line 17: @@
 [[file:T--TMMU-China--aimP.jpeg]]
-Then we inserted this gene to plasmid pSB1C3. We transformed this plasmid plays (one contains gene AimP) into strain DH5α (E. coli). Then we picked some colonies for cultivation and confirmed the transformation result by PCR. From the result of electrophoresis, we confirmed the transformation of AimP was success.
+Then we inserted this gene to plasmid pSB1C3. We transformed this plasmid plays (one contains gene AimP) into strain DH5α (<i>E. coli</i>). Then we picked some colonies for cultivation and confirmed the transformation result by PCR. From the result of electrophoresis, we confirmed the transformation of AimP was success.
 [[file:T--TMMU-China--aimp4.jpeg]]
@@ Line 24: / Line 24: @@
 ===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.
+The <i>B.subtilis</i> 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.
 [[file: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.
+By combining the expression of AimR and AimP components, we want to develop a synthetic QS system in <i>B.subtilis</i> for target gene autoinhibition.
 [[file: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.
+A synthetic communication pathway between <i>B.subtilis</i> strains by co-culturing AimP-producing “sender” cells with AimR-sensing “receiver” cells to inhibit gene expression was also designed.
 [[file:AimSR.jpeg]]