Because we found that FliC code includes restriction enzyme(EcoRI,SpeI,PstI) which is used in Biobrick, we divided into plasmid of FliC and one of signal, and aimed the double transformation.
- We regulate the expression of His-tagged FliC by lux promoter. Namely, if LuxR is expressed, bacteria can express FliC via Quorum Seinsing.
- Because the Quorum Sensing device is on ColE1-type vector, we need to export the FliC unit into the plasmids with compatible origins such as p15A.
First attempt to import FliC-His generator into p15A vector (see Fig. 2).
- Communication units are on ColE1 type plasmids. To make this stickly FliC construct compatible with communication circuits, this is an absolute necessity.....
- Not yet (as of 10/26/2007). The first ligation sucked. Cloning is still underway.
- Making Biobrick version of Flic-His and other stickly-FliCs. Unfortunately, Many forbidden sites for Biobrick production throughout the FliC gene. We are now eliminating them one by one by site-directed mutagenesis (ExSite PCR method).