Team:HokkaidoU Japan/Shuffling Kit/Future Work


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Maestro E. coli

Shuffling Kit

Maestro E. coli Future Work

To make Promoter Selector better kit

To make Promoter Selector better, we are going to place pigment producing construct between two PstI sites. After you confirm the best promoter, you can remove the followed constructs by digesting with PstI.

fig.1 How to make our kit better.

Application of RBS Selector

We can get a lot of operons by RBS Selector. RBS Selector provides 16 operons in case of 2 CDS, 64 operons in case of 3 CDS. What can we do using these number of operons? We expect industrial applications of RBS Selector as future works.

In the case of producing valuable molecules inside the cell, it is important to select the RBS which results the largest yield. For example PHB (polyhydroxybutyrate) ,3 enzymes synthesize PHB from substrate. PHB is one of the eco-friendly, biodegradable plastic. You can learn more about PHB at iGEM HokkaidoU_Japan 2012 wiki. In the production of PHB, each CDS has the optimum RBS. Our RS can be used for selecting the best combination of RBSs. It is possible to use RBS Selector when producing medicine or vaccine etc.

fig.2 P(3HB) synthesis pathway in R. eutropha.

Select Promoter & RBS

To select proteins expressions in wider range, you can use Promoter Selector and RBS Selector at the same time! The combination number is 320 patterns!

fig.3 Making various combination patarns.

Besides, you can create promoter and terminator by yourself and assemble them with our RBS Selector and GGA VECTOR (K1084301) by Golden Gate Assembly!