Difference between revisions of "Part:BBa K515102"
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| − | <p><i>Figure 1: Probability density function of bacterial number at observed velocities. PA2652 cells are more likely to be found at velocity of Rising concentrations of serine were tested. a) 0 mM control - circular colony b) 0.01 mM - circular colony e) 0.1 mM - circular colony d) 1 mM - possible eliptical colony c) 10 mM - eliptical colony f) 100 mM - eliptical colony away from the attractant. Data collected by Imperial iGEM 2011.</i></p> | + | <p><i>Figure 1: Probability density function of bacterial number at observed velocities. PA2652 cells exposed to malate are more than 90% likely to be moving at just over 2 μm/s. PA2652 cells that were exposed to serine were have likely to be found at velocity of Rising concentrations of serine were tested. a) 0 mM control - circular colony b) 0.01 mM - circular colony e) 0.1 mM - circular colony d) 1 mM - possible eliptical colony c) 10 mM - eliptical colony f) 100 mM - eliptical colony away from the attractant. Data collected by Imperial iGEM 2011.</i></p> |
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<h2>References</h2> | <h2>References</h2> | ||
<p>[1] Alvarez-Ortega C and Harwood CS (2007) Identification of malate chemoreceptor in <i>Pseudomonas aeruginosa</i> by screening for chemotaxis defects in an energy taxis-deficient mutant. <i>Applied and Environmental Microbiology</i> <b>73</b> 7793-7795.</p> | <p>[1] Alvarez-Ortega C and Harwood CS (2007) Identification of malate chemoreceptor in <i>Pseudomonas aeruginosa</i> by screening for chemotaxis defects in an energy taxis-deficient mutant. <i>Applied and Environmental Microbiology</i> <b>73</b> 7793-7795.</p> | ||
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Revision as of 01:07, 22 September 2011
J23100 promoter - PA2652
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 710
Illegal NgoMIV site found at 812
Illegal AgeI site found at 118 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1019
This BioBrick has been sequence verified.
Background
Malate responding chemoreceptor originally found in Pseudomonas aeruginosa PA01[1] is a codon optimised translational subunit. This subunit BBa_K515002 contains optimised insulator and RBS sequence, its expression is under the control of constitutive promoter BBa_J23100. Device is used as an additional chemoreceptor for endogenous chemotaxis mechanism of E. coli. Device responds to L (-) malic acid, linear formula (HO2CCH2CH(OH)CO2H).
Device contains 15 bp insulator seuence, which ensures tunability of expression through easy switching of promoters. In addition, insulator sequence allows the translation initiation rate (TIR) of the ribosome binding site (RBS) to remain the same, when the promoter is replaced.
Device is compatible for motile strains of E. coli. It has been tested in E. coli DH5α strain, inserted in the vector backbone pSB1C3.
Experimental Data
Behavioral analysis of E. coli cells was used to identify functioning of this device. The analysis is based on the transient difference in velocities of bacteria capable of sensing chemoattractant, when exposed to attractant which is in comparison to control cells, that are unable to sense L (-) malic acid.
Figure 1: Probability density function of bacterial number at observed velocities. PA2652 cells exposed to malate are more than 90% likely to be moving at just over 2 μm/s. PA2652 cells that were exposed to serine were have likely to be found at velocity of Rising concentrations of serine were tested. a) 0 mM control - circular colony b) 0.01 mM - circular colony e) 0.1 mM - circular colony d) 1 mM - possible eliptical colony c) 10 mM - eliptical colony f) 100 mM - eliptical colony away from the attractant. Data collected by Imperial iGEM 2011.
References
[1] Alvarez-Ortega C and Harwood CS (2007) Identification of malate chemoreceptor in Pseudomonas aeruginosa by screening for chemotaxis defects in an energy taxis-deficient mutant. Applied and Environmental Microbiology 73 7793-7795.