Difference between revisions of "Part:BBa K2656009"
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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 optimized values based on each RBS tested. | ||
| − | + | [[File:https://static.igem.org/mediawiki/2018/b/b1/T--Valencia_UPV--optimization_exp1_RBS_graphUPV2018.png|900px|thumb|none|alt=RBS experiment 1.|Figure 1. RBS expression experiment with K2656008, K2656009 and K2656012]] | |
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| − | + | [[File:T--Valencia_UPV--optimization_exp2sfGFP_all_graphUPV2018.png|900px|thumb|none|alt=RBS experiment 2.|Figure 2. RBS expression experiment with K2656008 and K2656012]] | |
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|colspan=4|Table 1. Optimized parameters for the BBa_K2656009 RBS. | |colspan=4|Table 1. Optimized parameters for the BBa_K2656009 RBS. | ||
Revision as of 21:07, 14 October 2018
Strong Ribosome Binding Site B0030
BBa_K2656009 is the BBa_B0030 RBS 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 two comparative RBS expression experiments with composite parts BBa_K2656102, BBa_K2656103 and [1],BBa_K2656100, [2],BBa_K2656104 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]
| Table 1. Optimized parameters for the BBa_K2656009 RBS. | |||
| Parameter | Value | ||
| Translation rate p | p = 0.2059 min-1 | ||
| Dilution rate μ | μ = 0.01492 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 | 1 | ||
| p parameter ratio (pRBS/pref) | 1 | ||
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]