Difference between revisions of "Part:BBa K2656011"

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−	By using this [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol], we have obtained the parameters to valide our  [http://2018.igem.org/Team:Valencia_UPV/Modeling#models constitutive model]and rationale choose its optimized values based on each RBS tested.	+	By using this [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol], we have obtained the parameters to valide our  [http://2018.igem.org/Team:Valencia_UPV/Modeling#models constitutive model]and rationale choose its [http://2018.igem.org/Team:Valencia_UPV/Modeling#optimization optimization values] based on each RBS tested.
	[[File:T--Valencia_UPV--optimization_exp1_RBS_graphUPV2018.png|900px|thumb|none|alt=RBS experiment 1.|Figure 1. RBS expression experiment with K2656009, K26560010 and K2656011 RBS basic parts]]		[[File:T--Valencia_UPV--optimization_exp1_RBS_graphUPV2018.png|900px|thumb|none|alt=RBS experiment 1.|Figure 1. RBS expression experiment with K2656009, K26560010 and K2656011 RBS basic parts]]

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Revision as of 21:58, 14 October 2018

Medium Ribosome Binding Site B0034

BBa_K2656011 is the original RBS B0034 standardized into the Golden Braid assembly method. Thus, it is strong ribosome binding site compatible both with the BioBrick and Golden Gate grammar.

Using the [http://2018.igem.org/Team:Valencia_UPV/Protocols Golden Gate assembly protocol], it can be combined with other GB adapted basic parts, such as the ones from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Valencia UPV IGEM 2018 Printeria Collection], to assemble transcriptional units in a single one-pot reaction.

Characterization of the this part was performed with the transcriptional unit BBa_K2656101 , which was used in a comparative RBS expression experiment with composite parts BBa_K2656101 and BBa_K2656102. They all were assembled in a Golden Braid alpha1 plasmid using the same promoter, CDS and terminator:

• Promoter BBa_K2656004: the J23106 promoter in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]
• CDS BBa_K2656013: the BBa_K2656009 sfGFP sequence in its Golden Braid standardized version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]
• Terminator BBa_K2656026: the B0015 transcriptional terminator in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]

By using this [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol], we have obtained the parameters to valide our [http://2018.igem.org/Team:Valencia_UPV/Modeling#models constitutive model]and rationale choose its [http://2018.igem.org/Team:Valencia_UPV/Modeling#optimization optimization values] based on each RBS tested.

Table 1. Optimized parameters for the BBa_K2656009 RBS.
Parameter	Value
Translation rate p	p = 0.082 min-1
Dilution rate μ	μ = 0.01631 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 (GB BBa_B0032 RBS) relative strength and p ratio.
Parameter	Value
Relative strength	0.371
p parameter ratio (pRBS/pref)	0.398

Sequence and Features