Difference between revisions of "Part:BBa K1031923"
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<p style="text-align:center"><b>Fig 2</b> Performance of different XylB Adptor with different culture condition. Final concentrations of inducer were 300 uM. BAL: benzyl alcohol; 2-MeBAL: 2-methylbenzyl alcohol; Coculture: M9 minimal media containing the aromatics mentioned above was processed by E. coli transformed with Adaptor XylB and E. coli transformed with XylC simultaneously; Double Trans.: double transformation; Dashed box refers to performance of XylS Biosensor equipped with Adaptor XylC and K1031923.  
 
<p style="text-align:center"><b>Fig 2</b> Performance of different XylB Adptor with different culture condition. Final concentrations of inducer were 300 uM. BAL: benzyl alcohol; 2-MeBAL: 2-methylbenzyl alcohol; Coculture: M9 minimal media containing the aromatics mentioned above was processed by E. coli transformed with Adaptor XylB and E. coli transformed with XylC simultaneously; Double Trans.: double transformation; Dashed box refers to performance of XylS Biosensor equipped with Adaptor XylC and K1031923.  
 
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<p style="text-align:center"><b>Fig 3</b> Characterization and comparison of biosensor XylS with or without Adaptor XylB and XylC. Final concentrations of inducers were 0, 0.3, 1, 3, 10, 30, 100, 300 uM in the characterization assay. <b>(a)</b> Dose-response curve for benzyl alcohol (BAL); <b>(b)</b> Dose-response curve for 2-methylbenzyl alcohol (2-MeBAL); <b>(c)</b> Dose-response curve for benzoate (BzO); (d) Dose-response curve for benzoate (2-MeBzO).
 
<p style="text-align:center"><b>Fig 3</b> Characterization and comparison of biosensor XylS with or without Adaptor XylB and XylC. Final concentrations of inducers were 0, 0.3, 1, 3, 10, 30, 100, 300 uM in the characterization assay. <b>(a)</b> Dose-response curve for benzyl alcohol (BAL); <b>(b)</b> Dose-response curve for 2-methylbenzyl alcohol (2-MeBAL); <b>(c)</b> Dose-response curve for benzoate (BzO); (d) Dose-response curve for benzoate (2-MeBzO).
 
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Revision as of 05:44, 27 October 2013

J23109-XylB-Terminator

For detailed information concerning XylB, please visit 2013 Peking iGEM Adaptors


Characterization

We achieved the concept of Adaptor by connecting XylC with XylS, and we want to further confirm whether other degradation enzymes could be added. So we construct XylB adaptor circuit adopting J23109 as constitutive promoter and B0015 as terminator (Fig 1)

Fig 1 Construction of adaptor circuit Pc/J23109-XylB. The orange arrowhead refers to constitutive Pc promoter. RBS is shown in green oval. The square in dark red stands for terminator B0015.


Sequence and Features

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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 364


Data shown

A set of culturing modes were tried respectively to optimize the performance of Adaptor XylB. They were listed in Table 1.


For part K1031923 adopting constitutive Pc promoter J23109, only the third protocol works. Induction ratio is shown in Fig 2when exposed to BAL and 2-MeBAL. The dashed box refers to the performance of XylS Biosensor equipped with XylC and J23109-XylB Adptors.

Fig 2 Performance of different XylB Adptor with different culture condition. Final concentrations of inducer were 300 uM. BAL: benzyl alcohol; 2-MeBAL: 2-methylbenzyl alcohol; Coculture: M9 minimal media containing the aromatics mentioned above was processed by E. coli transformed with Adaptor XylB and E. coli transformed with XylC simultaneously; Double Trans.: double transformation; Dashed box refers to performance of XylS Biosensor equipped with Adaptor XylC and K1031923.



Dose-response Curves were plotted to further describe response performance when exposed to four inducers respectively at different concentrations. Each strain is marked in different colors. The curves in blue represents Biosensor equipped with K1031923.

Fig 3 Characterization and comparison of biosensor XylS with or without Adaptor XylB and XylC. Final concentrations of inducers were 0, 0.3, 1, 3, 10, 30, 100, 300 uM in the characterization assay. (a) Dose-response curve for benzyl alcohol (BAL); (b) Dose-response curve for 2-methylbenzyl alcohol (2-MeBAL); (c) Dose-response curve for benzoate (BzO); (d) Dose-response curve for benzoate (2-MeBzO).