Difference between revisions of "Part:BBa K1031923"
 
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<img src="https://static.igem.org/mediawiki/2013/f/fc/Peking2013_part_109-XylB.png" style="width:450px; margin-left:220px" />
 
<img src="https://static.igem.org/mediawiki/2013/f/fc/Peking2013_part_109-XylB.png" style="width:450px; margin-left:220px" />
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<p style="text-align:center"><b>Fig 1</b> Construction of adaptor circuit <i>Pc</i>/J23109-XylB. The orange arrowhead refers to constitutive <i>Pc</i> promoter. RBS is shown in green oval. The square in dark red stands for terminator B0015.
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<p style="text-align:center"><b>Fig 1</b> Construction of adaptor circuit J23109-XylB. The orange arrowhead refers to constitutive <i>Pc</i> promoter. RBS(Ribosome Binding Site) is shown in green oval. The square in dark red stands for terminator B0015.
 
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== '''Data shown''' ==
 
== '''Data shown''' ==
  
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A set of culturing conditions were tried respectively to optimize the performance of K1031923. They were listed in '''Table 1''', marked in blue.
  
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('''Fig 2''').  
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<img src="https://static.igem.org/mediawiki/2013/thumb/c/cf/Peking2013_part_109-XylB_Table.png/800px-Peking2013_part_109-XylB_Table.png" style="width:600; margin-left:70px" />
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For Biobrick K1031923 adopting constitutive ''Pc'' promoter J23109, only the third protocol works. Induction ratio is shown in '''Fig 2'''when exposed to BAL and 2-MeBAL. The dashed box refers to the performance of XylS Biosensor equipped with XylC and J23109-XylB Adptors.
 
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<img src="https://static.igem.org/mediawiki/2013/thumb/c/cc/Peking2013_part_109-XylB_library.png/800px-Peking2013_part_109-XylB_library.png"
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<p style="text-align:center"><b>Fig 2</b> Schematic diagrams for the plasmid circuits used as Adaptor: NahF and the Sensor: NahR. A constitutive promoter library for the expression of NahF was constructed to obtain the most appropriate expression level of NahF enzyme in E.coli. The number of the Standard Biological constitutive promoter Parts used in this study and its initiation strength is listed in the left portion of the figure. Promoters are presented in orange, RBS in light green, coding sequence in dark cyan, and terminators in dark red.
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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. Abbreviations used in this figure: 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. The Dashed box refers to performance of XylS Biosensor equipped with Adaptor XylC and K1031923.  
 
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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 performance of XylS Biosensor equipped with 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); <b>(d)</b> Dose-response curve for benzoate (2-MeBzO).
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Latest revision as of 16:53, 28 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 J23109-XylB. The orange arrowhead refers to constitutive Pc promoter. RBS(Ribosome Binding Site) 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 conditions were tried respectively to optimize the performance of K1031923. They were listed in Table 1, marked in blue.


For Biobrick 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. Abbreviations used in this figure: 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. The 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 performance of XylS 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).