Difference between revisions of "Part:BBa K515102"
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<p><b>This BioBrick has been sequence verified.</b>
 
<p><b>This BioBrick has been sequence verified.</b>
 
<h2>Background</h2>
 
<h2>Background</h2>
−
<p>Malate responding chemoreceptor originally found in <i>Pseudomonas aeruginosa</i> PA01<sup>[1]</sup> is a codon optimised translational subunit. This subunit <a href="https://parts.igem.org/Part:BBa_K515002">BBa_K515002</a> contains optimised insulator and RBS sequence, its expression is under the control of constitutive promoter <a href="https://parts.igem.org/Part:BBa_J23100">BBa_J23100</a>. Device is used as an additional chemoreceptor for endogenous chemotaxis mechanism of <i>E. coli</i>. Device responds to L (-) Malic acid, linear formula (HO2CCH2CH(OH)CO2H).</p>
+
<p>Malate responding chemoreceptor originally found in <i>Pseudomonas aeruginosa</i> PA01<sup>[1]</sup> is a codon optimised translational subunit. This subunit <a href="https://parts.igem.org/Part:BBa_K515002">BBa_K515002</a> contains optimised insulator and RBS sequence, its expression is under the control of constitutive promoter <a href="https://parts.igem.org/Part:BBa_J23100">BBa_J23100</a>. Device is used as an additional chemoreceptor for endogenous chemotaxis mechanism of <i>E. coli</i>. Device responds to L (-) malic acid, linear formula (HO2CCH2CH(OH)CO2H).</p>
−
<p> 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 translational subunit to remain the same, when the promoter is replaced.</p>
+
<p>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.</p>
−
 
+
<p>Device is compatible for motile strains of <i>E. coli</i>. It has been tested in <i>E. coli</i> DH5α strain, inserted in the vector backbone <a href="https://parts.igem.org/Part:pSB1C3">pSB1C3</a>.</p>
−
<b>Compatibility</b>
+
<h2>Experimental Data</h2>
−
<p>Device has been tested in <i>E. coli DH5α</i> strain, inserted in vector backbone <a href="https://parts.igem.org/Part:pSB1C3">pSB1C3</a>. Note that device is only functional in motile strains of <i>E. coli</i>.</p>
+
<p>Behavioral analysis of <i>E. coli</i> 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 in comparison to control, which is unable to sense L (-) malic acid.</p>
 +
<div class="imgbox" style="width:920px;" >
 +
<img class="border" src="https://static.igem.org/mediawiki/parts/2/26/ICL_PA2652_probability_density_function.png" width="900px" />
 +
<p><i>Figure 1: 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>
 +
</div>
 
<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>
 
</html>
 
</html>

Revision as of 23:39, 21 September 2011

J23100 promoter - PA2652


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 710
    Illegal NgoMIV site found at 812
    Illegal AgeI site found at 118
  • 1000
    INCOMPATIBLE 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 in comparison to control, which is unable to sense L (-) malic acid.

Figure 1: 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.