Difference between revisions of "Part:BBa I13453"

Revision as of 16:17, 10 May 2013

Pbad promoter

PBad promoter from I0500 without AraC.

Usage and Biology

Has been used as a second promoter in a system containing BBa_I0500 (PBad+AraC). In this system, it showed behavior qualitatively indistinguishable from the BBa_I0500 copy of PBad. Has not been tested independent of AraC. A second part, BBa_I13458, should allow decoupling of PBad and AraC.

See this OpenWetWare article on [http://openwetware.org/wiki/Titratable_control_of_pBAD_and_lac_promoters_in_individual_E._coli_cells#pBAD_promotersOpenWetWare pBAD and lac promoters] for additional usage and biology information

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 125
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 65
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Characterization

For the 2009 iGEM competition, British_Columbia characterized BBa_I13453 in the context of a pBAD promoter family. For the results of this characterization, see here.

For the 2010 iGEM competition, Tec-Monterrey characterized BBa_I13453 again, using a construct of GFP reporter after the pBad promoter. The transfer function was modeled with a Hill equation.

Figure 1. Transfer function of BBa_I13453. Points represent individual measurements. The line is of a Hill equation fitted to our data.

(d[GFP]/dt)/OD₆₀₀ = C+A*Xⁿ/(Xⁿ+Kⁿ)

Experiment	Characteristic	Value
Transfer Function	Basal rate (C)	1 d[GFP]/dt
	Gain (A)	36 d[GFP]/dt
	Hill coefficient (n)	2.16
	Switch Point (K)	2.8 [ara] (µM)

MetA Knockout Complementation by inducing arabinose promoter by British Columbia iGEM 2012

This part consists of an arabinose promoter, a strong RBS, and the MetA coding gene. It is able to complement a metA knockout, and the growth rate appears to be proportional to the amount of arabinose that is added.

To generate this graph we cultured an E. coli MetA knockout transformed with this part in M9 minimal media with the indicated concentrations of arabinose, and measured the OD600 every 15 minutes.

TrpA Knockout Complementation by inducing arabinose promoter by British Columbia iGEM 2012

This part consists of an arabinose promoter, a strong RBS, and the TrpA coding gene. It is able to complement a metA knockout, and the growth rate appears to be proportional to the amount of arabinose that is added.

To generate this graph we cultured an E. coli TrpA knockout transformed with this part in M9 minimal media with the indicated concentrations of arabinose, and measured the OD600 every 15 minutes. See BBa_K804009 for negative controls for this part as well as linked fluorescent data. Not understood

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-1005123605 http://manualiguj.pp.ua/ http://manualiguj.pp.ua/tfhm1.html http://instructionsqsa.pp.ua/kml2.html  http://documentsjgq.pp.ua/bmy2.html http://documents4xm7.pp.ua/  http://manualip2cj.pp.ua/  http://documentspacs.pp.ua/ http://documents140.pp.ua/coyx1.html http://manualip2cj.pp.ua/ies1.html http://documents140.pp.ua/  http://manualip2cj.pp.ua/vnauw2.html http://documentsjgq.pp.ua/kuelta1.html    http://documentsjgq.pp.ua/      http://instructionsqsa.pp.ua/cfco1.html         http://manualiguj.pp.ua/evr2.html    http://documents140.pp.ua/rlivwk2.html
+__NOTOC__
+<partinfo>BBa_I13453 short</partinfo>
+PBad promoter from I0500 without AraC.
+===Usage and Biology===
+Has been used as a second promoter in a system containing <partinfo>I0500</partinfo> (PBad+AraC). In this system, it showed behavior qualitatively indistinguishable from the <partinfo>I0500</partinfo> copy of PBad. Has not been tested independent of AraC. A second part, <partinfo>I13458</partinfo>, should allow decoupling of PBad and AraC.
+See this OpenWetWare article on [http://openwetware.org/wiki/Titratable_control_of_pBAD_and_lac_promoters_in_individual_E._coli_cells#pBAD_promotersOpenWetWare pBAD and lac promoters] for additional usage and biology information
+<span class='h3bb'>Sequence and Features</span>
+<partinfo>BBa_I13453 SequenceAndFeatures</partinfo>
+===Characterization===
+For the 2009 iGEM competition, British_Columbia characterized <partinfo>I13453</partinfo> in the context of a [https://parts.igem.org/PBAD_Promoter_Family pBAD promoter family]. For the results of this characterization, see [[Part:BBa_I13453:British_Columbia|here]].
+For the 2010 iGEM competition, Tec-Monterrey characterized <partinfo>I13453</partinfo> again, using a construct of GFP reporter after the pBad promoter. The transfer function was modeled with a Hill equation.
+<!-- -->
+[[Image:pbadwt.png|center|frame|Figure 1. Transfer function of <partinfo>BBa_I13453</partinfo>. Points represent individual measurements. The line is of a Hill equation fitted to our data.]]
+<center>
+(d[GFP]/dt)/OD<sub>600</sub> = C+A*X<sup>n</sup>/(X<sup>n</sup>+K<sup>n</sup>)
+{|{{Table}}
+!Experiment
+!Characteristic
+!Value
+|-
+|rowspan="4"|[[#Transfer Function|'''Transfer Function''']]
+|''Basal rate (C)''
+|1 d[GFP]/dt
+|-
+|''Gain (A)''
+|36 d[GFP]/dt
+|-
+|''Hill coefficient (n)''
+|2.16
+|-
+|''Switch Point (K)''
+|2.8 [ara] (µM)
+|-
+|}
+</center>
+<!-- Uncomment this to enable Functional Parameter display
+===Functional Parameters===
+<partinfo>BBa_I13453 parameters</partinfo>
+<!-- -->
+===MetA Knockout Complementation by inducing arabinose promoter by British Columbia iGEM 2012===
+This part consists of an arabinose promoter, a strong RBS, and the MetA coding gene. It is able to complement a metA knockout, and the growth rate appears to be proportional to the amount of arabinose that is added.
+<p align=center> https://static.igem.org/mediawiki/parts/thumb/d/d3/MetA_complementation_no_fluor.png/800px-MetA_complementation_no_fluor.png</p>
+To generate this graph we cultured an E. coli MetA knockout transformed with this part in M9 minimal media with the indicated concentrations of arabinose, and measured the OD600 every 15 minutes.
+===TrpA Knockout Complementation by inducing arabinose promoter by British Columbia iGEM 2012===
+This part consists of an arabinose promoter, a strong RBS, and the TrpA coding gene. It is able to complement a metA knockout, and the growth rate appears to be proportional to the amount of arabinose that is added.
+https://static.igem.org/mediawiki/parts/thumb/0/04/TrpA_Complementation_Test_No_fluor.png/800px-TrpA_Complementation_Test_No_fluor.png
+To generate this graph we cultured an E. coli TrpA knockout transformed with this part in M9 minimal media with the indicated concentrations of arabinose, and measured the OD600 every 15 minutes. See <partinfo>BBa_K804009 for negative controls for this part as well as linked fluorescent data.