RBS

Part:J61101

Designed by: John Anderson   Group: Arkin Lab   (2007-01-28)

Contribution

Group: Valencia_UPV iGEM 2018
Author: Adrián Requena Gutiérrez, Carolina Ropero
Summary: We adapted the part to be able to assemble transcriptional units with the Golden Gate assembly method
Documentation: In order to create our complete part collection of parts compatible with the Golden Gate assembly method, we made the part BBa_K2656012 which is this part adapted to the Golden Gate technology.

Characterization of the this part was performed with the transcriptional unit BBa_K2656104, which was used in a comparative RBS expression experiment with composite parts BBa_K265610 and BBa_K2656101. They all were assembled in a Golden Braid alpha1 plasmid using the same promoter, CDS and terminator.

By using this experimental protocol, we have obtained the parameters to valide our constitutive modeland rationale choose its optimized values based on each RBS.

RBS experiment.
Figure 1. RBS expression experiment with K2656008, K2656009 and K2656012 RBS basic pieces
RBS experiment.
Figure 2. RBS expression experiment with K2656008 and K2656012 RBS basic pieces
Table 1. Optimized parameters for the TU with BBa_K2656008 RBS.
Parameter Value
Translation rate p p = 0.02889 min-1
Dilution rate μ μ = 0.0118 min-1

We have also calculated the relative force between the different RBS, taking BBa_K2656009 strong RBS as a reference. Likewise, a ratio between p parameters of the different RBS parts and p parameter of the reference RBS has been calculated.

Table 2. BBa_K2656008 relative strength and p ratio.
Parameter Value
Relative strength 0.032
p parameter ratio (pRBS/pref) 0.032
[edit]
Categories
//rbs/prokaryote/constitutive/anderson
//ribosome/prokaryote/ecoli
//chassis/prokaryote/ecoli
//direction/forward
//regulation/constitutive
Parameters
None