Team:TU Delft/project/rbs characterization

From 2010.igem.org

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=RBS Characterization=
=RBS Characterization=
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For our RBS characterization project, 5 different RBS sequences from the [http://partsregistry.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Anderson Anderson RBS family] where placed in front of GFP and measured over 18 hours using a Gen5 fluorenscence and absorbance plate reader
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[[Image:26_07_2010_Rbs.png]]
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Strength was calculated by taking the mean of the ratio between the expression of known RBS ([http://partsregistry.org/Part:BBa_B0032 B0032]) and expression of Anderson RBS over some time. Expression being the measured fluorescence divided by measured biomass (absorbance, OD).
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The measurements where taken from the part of the curve where optimal growth can be assumed, so from 0:40 until 2:30
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The result is a simple characterization of the Anderson RBS sequences in relation to the known RBSes.
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{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
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|'''RBS'''
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|'''Strength'''
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|-
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|[http://partsregistry.org/Part:BBa_J61100 J61100]
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|0.047513
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|-
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|[http://partsregistry.org/Part:BBa_J61101 J61101]
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|0.119831
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|-
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|[http://partsregistry.org/Part:BBa_J61107 J61107]
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|0.065454
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|-
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|[http://partsregistry.org/Part:BBa_J61117 J61117]
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|0.038518
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|-
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|[http://partsregistry.org/Part:BBa_J61127 J61127]
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|0.087334
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|-
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|[http://partsregistry.org/Part:BBa_B0032 B0032]
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|0.300000
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|}
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We've also looked at mRNA folded shapes using mfold to see if there was a common pattern in the Anderson RBS shapes. Unfortunately, this doesn't seem to be the case, as all the tried RBS sequences had very different mfold shapes.

Revision as of 12:04, 2 August 2010

RBS Characterization

For our RBS characterization project, 5 different RBS sequences from the Anderson RBS family where placed in front of GFP and measured over 18 hours using a Gen5 fluorenscence and absorbance plate reader 26 07 2010 Rbs.png

Strength was calculated by taking the mean of the ratio between the expression of known RBS (B0032) and expression of Anderson RBS over some time. Expression being the measured fluorescence divided by measured biomass (absorbance, OD).

The measurements where taken from the part of the curve where optimal growth can be assumed, so from 0:40 until 2:30 The result is a simple characterization of the Anderson RBS sequences in relation to the known RBSes.

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. Unfortunately, this doesn't seem to be the case, as all the tried RBS sequences had very different mfold shapes.