Difference between revisions of "Part:BBa K1031621"
Line 1: | Line 1: | ||
− | |||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1031621 short</partinfo> | <partinfo>BBa_K1031621 short</partinfo> | ||
− | + | <!-- --> | |
+ | |||
+ | |||
+ | == '''Characterization''' == | ||
+ | |||
+ | |||
+ | In order to fine-tune the performance of NahF adapter for NahR biosensor, we constructed a library of ''Pc'' constitutive promoters at different intensity. BBa_K1031621 is composed of three elements, the constitutive ''Pc'' promoter J23105[https://parts.igem.org/Part:BBa_J23105], coding sequence of NahF and terminator B0015[https://parts.igem.org/Part:BBa_B0015]. ('''Fig 1''') | ||
+ | |||
+ | <html> | ||
+ | <img src="https://static.igem.org/mediawiki/igem.org/2/2a/Peking2013_part_105-NahF.png", width=600px; /> | ||
+ | </html> | ||
+ | |||
+ | '''Fig 1''' Construction of adapter circuit ''Pc''/J23105-NahF | ||
+ | |||
+ | |||
+ | == '''Sequence and Features''' == | ||
− | |||
− | |||
<!-- --> | <!-- --> | ||
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K1031621 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1031621 SequenceAndFeatures</partinfo> | ||
+ | |||
+ | |||
+ | |||
+ | == '''Data shown''' == | ||
+ | |||
+ | |||
+ | It is necessary to fine-tune the expression level of NahF in ''E.coli''. We built a library of constitutive promoters for tuning the expression of NahF, and NahR biosensor was used to detect the possible salicylates transformed from salicylaldehydes ('''Fig 2a'''). The dashed box refers to data for ''Pm''/J23105-XylS biosensor circuit ('''Fig 2b''') | ||
+ | |||
+ | <html> | ||
+ | <img src="https://static.igem.org/mediawiki/igem.org/2/2a/Peking2013_part_105-NahF.png, width=600px; /> | ||
+ | </html> | ||
+ | |||
+ | '''Fig 2''' Construction and test results of NahR biosensor equipped with NahF adapter | ||
+ | '''a'''. 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.intensity | ||
+ | '''b'''. Horizontal axis stands for NahR biosensor equipped with NahF adapters adopting ''Pc'' promoters of different strength. The expression intensity of these constitutive promoters, J23113, J23109, J23114, J23105, J23106 is 21, 106, 256, 623, and 1185, respectively, according to the Part registry. Two kinds of aromatics, namely ... and ..., shown with different color intensities, were tested following Test Protocol 1 [1]. Vertical axis represents the ON/OFF induction ratio. The dashed box refers to performance for ''Pc''/J23105-NahF adapter circuit | ||
Revision as of 18:13, 24 September 2013
J23105-NahF-Terminator
Characterization
In order to fine-tune the performance of NahF adapter for NahR biosensor, we constructed a library of Pc constitutive promoters at different intensity. BBa_K1031621 is composed of three elements, the constitutive Pc promoter J23105[1], coding sequence of NahF and terminator B0015[2]. (Fig 1)
Fig 1 Construction of adapter circuit Pc/J23105-NahF
Sequence and Features
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 906
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1144
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1042
Illegal BsaI.rc site found at 1196
Data shown
It is necessary to fine-tune the expression level of NahF in E.coli. We built a library of constitutive promoters for tuning the expression of NahF, and NahR biosensor was used to detect the possible salicylates transformed from salicylaldehydes (Fig 2a). The dashed box refers to data for Pm/J23105-XylS biosensor circuit (Fig 2b)