Tokyo/Works/Hybrid promoter

From 2007.igem.org

< Tokyo/Works
Revision as of 17:48, 24 October 2007 by Hiro (Talk | contribs)


Works top  0. Hybrid promoter  1. Formulation  2. Assay1  3. Simulation  4. Assay2  5. Future works

Purpose

Fig.1: Genetic Circuit

To realize our model, expression of one part of the genes should be regulated by two factors. Therefore, the promoter upstream of the idlers genetic part was designed to sense TWO INPUTS, activated by AHL and repressed by LacI.


Design

Based on the AHL regulated BioBrick part R0062 and the previous work on LacI regulated promoter (ref. 1), we designed a new promoter regulated by AHL and LacI. Since this promoter is a kind of a "HYBRID" of the two, we call it HYBRID PROMOTER.

Fig.2: Genetic Circuit


Compared with existing BioBrick hybrid promoter R0065, this promoter is different in that its sequence is much shorter with lux box and -35 region overlapping, and that it is repressed by LacI, not cI.
Also, the output is clearly detected as shown in Fig. 2.

References
-Biobrick R0062
-Lutz, R., and H. Bujard. 1997. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I-1-I-2 regulatory elements. Nucleic Acids Res. 25:1203–1210.

Experiments

Fig.3: The result of assay0

The characteristics of this hybrid promoter (Lux-lac hybrid promoter) have been determined though Assay 0. The results are shown in Fig.3.

 


Works top << Hybrid promoter >> Next(Formulation)