Team:TU Delft/Project/rbs-characterization


Revision as of 16:53, 19 October 2010 by Kschipper (Talk | contribs)

Ribosome Binding Site Characterization

TUDelft RBS.png

Part of our project is to measure the effect of various ribosome binding site (RBS) sequences. A RBS sequence is a specific mRNA sequence that folds in such a way that it attracts the ribosome. This ribosome binds to the mRNA molecule and starts translation of the mRNA into protein. Accurate information on how well this binding occurs (The binding strength) can be very useful when designing new biological systems.

We took five different RBS sequences from the Anderson RBS family (J61100, J61101, J61107, J61117, J61127). The standard RBS B0032 was used as a reference for our characterization.

All these RBS sequences were placed in front of the standard GFP coding sequence, so expression could be measured. The Biobricks generated in order to perform the experiments were: K398500, K398501, K398502, K398503, K398504. The general map of the construction is shown below, where the RBS is displayed in fucsia.

Feature Function
AmpR Ampicillin resistance
B0015 Transcriptional (double) terminator
B0062 Transcriptional terminator
E0040 GFP
G00000 Standard prefix
G00001 Standard suffix
G00100 VF2 primer binding site
G00101 VR primer binding site
J61100 RBS Anderson family
J23100 Promoter