Difference between revisions of "Part:BBa K2666008"

Latest revision as of 08:15, 15 October 2018

RFP with B. subtilis specific strong promoter R0

I. Part BBa K2666001 : Function

The Montpellier iGEM team 2018 designed a construction that produce RFP in Bacillus subtilis (http://2018.igem.org/Team:Montpellier/Toolbox). For that, we used the promoter R0, a B.subtilis specific strong promoter [1]. Figure 1 illustrates the detailed design.

Figure 1 : Construct design. The sequence that encode RFP fusionned with a strong promoter R0.

The objective is to check the strength of several construction in E.coli, L.jensenii & B.subtilis.

II. Proof of function

Figure 2 : Quantity of E.coli that produce RFP by low cytometer. In gray, the control without plasmid. In pink, the E. coli population with the construction R0-RFP.

Figure 3 : Number of E.coli that produce RFP by low cytometer. A) E. coli population with the construction R0-RFP B) Control without plasmid.

III. Design Considerations

We added spacers to all of our constructions to unable easier use of the sequence and separation of the different genes of the sequences. We used two Terminators to our sequences :BBa_B0014 & BBa_B0015 to ensure the stopping of the transcription.

Reference:

[1] Guiziou, Sarah, et al. "A part toolbox to tune genetic expression in Bacillus subtilis." Nucleic acids research 44.15 (2016): 7495-7508.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 62
Illegal BamHI site found at 194
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
-==  LL-37 with L.jensenii specific strong promoter RpsU ==
+== RFP with B. subtilis specific strong promoter R0 ==
 === I. Part BBa K2666001 : Function ===
 The Montpellier iGEM team 2018 designed a construction that produce RFP in Bacillus subtilis (http://2018.igem.org/Team:Montpellier/Toolbox). For that, we used the promoter R0, a B.subtilis specific strong promoter [1]. Figure 1 illustrates the detailed design.
+[[File:T--Montpellier--R0-RFP design mtp.PNG|550px]]
 '''Figure 1''' : Construct design. The sequence that encode RFP fusionned with a strong promoter R0.
 The objective is to check the strength of several construction in E.coli, L.jensenii & B.subtilis.
 === II. Proof of function ===
+[[File:T--Montpellier--R0-RFP Cytometer mtp.PNG|350px]]
+'''Figure 2''' : Quantity of E.coli that produce RFP by low cytometer. In gray, the control without plasmid. In pink, the E. coli population with the construction R0-RFP.
+[[File:T--Montpellier--R0-RFP cytometer2 mtp.PNG|500px]]
+'''Figure 3''' : Number of E.coli that produce RFP by low cytometer. A) E. coli population with the construction R0-RFP B) Control without plasmid.
 === III. Design Considerations ===
 We added spacers to all of our constructions to unable easier use of the sequence and separation of the different genes of the sequences. We used two Terminators to our sequences :BBa_B0014 & BBa_B0015 to ensure the stopping of the transcription.
 == Reference: ==