Team:TU Delft/project/rbs characterization
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- | For our RBS characterization project, | + | For our RBS characterization project, five different RBS sequences from the [http://partsregistry.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Anderson Anderson RBS family] (J61100, J61101, J61107, J61117, J61127) and the standard RBS [http://partsregistry.org/Part:BBa_B0032 B0032] were placed in front of the GFP generator; the protein production was measured using fluorescence as main parameter. The experiments were carried out during 18 hours using a Gen5 fluorenscence and absorbance plate reader. The general map of the construction is shown below, where the RBS is displayed in fucsia. |
[[Image:RBS1.jpg|left|650px]] | [[Image:RBS1.jpg|left|650px]] | ||
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Revision as of 13:32, 10 August 2010
RBS Characterization
For our RBS characterization project, five different RBS sequences from the Anderson RBS family (J61100, J61101, J61107, J61117, J61127) and the standard RBS B0032 were placed in front of the GFP generator; the protein production was measured using fluorescence as main parameter. The experiments were carried out during 18 hours using a Gen5 fluorenscence and absorbance plate reader. 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 |
RBS | Strength |
J61100 | 0.047513 |
J61101 | 0.119831 |
J61107 | 0.065454 |
J61117 | 0.038518 |
J61127 | 0.087334 |
B0032 | 0.300000 |
We've also looked at mRNA folded shapes using mfold to see if there was a common pattern in the Anderson RBS shapes. This might have been usable in predicting RBS strength for the other untested Anderson RBS sequences. Unfortunately, this didn't seem to be the case, as all the tried RBS sequences had very different mfold shapes.