Reporter

Part:BBa_K2276007

Designed by: Tianying Chen   Group: iGEM17_SCU_China   (2017-10-24)
Revision as of 04:37, 31 October 2017 by Lilita (Talk | contribs) (→‎Measurement)

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a composite of TetR repressible promoter、special designed RBS and the coding sequence of mRFP

The part was designed based on the Part: BBa_I13521, while the RBS binding site is modified. The RBS is special designed for strainS whose last 9 nucleotides of the 16S rRNA are ACCTCCTTA. And it can be regard as a improvement for it has a high translation rate compared with Part: BBa_I13521.The promoter is TetR repressible promoter which is also used in the repressilator system. And the mRFP served as the reporter to help us measure the translation rate of the designed RBS.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 634
    Illegal AgeI site found at 746
  • 1000
    COMPATIBLE WITH RFC[1000]


Measurement

We constructed strains that contain Part:BBa K2276007 and measure the fluorescence intensity and OD600 changes of it. Due to the limit of the experiment condition, we measure the change at the first 8hours and 10~16 hours after inoculated separately. As what Fig 1 and Fig 2 show,Part: BBa_K2276007 has a much strong expression than Part: BBa_I13521.

fig.1The fluorescence intensity changes over time (0h~16h). P1 represent the strain containing part: BBa_K2276007. P2 represent the strain containing part: BBa_K2276008. P3 represent the strain containing part: BBa_K2276010. TetR represent the strain that only has tetR repressible promoter without RBS and protein coding sequence. B0034 represent the strain that containing Part: BBa_I13521.
fig.2 The OD600 changes over time(0~16 h).P1 represent the strain containing part: BBa_K2276007. P2 represent the strain containing part: BBa_K2276008. P3 represent the strain containing part: BBa_K2276010. TetR represent the strain that only has tetR repressible promoter without RBS and protein coding sequence. B0034 represent the strain that containing Part: BBa_I13521.

Source

BBa_I13521

References

[1] Borujeni A E, Salis H M. Translation Initiation is Controlled by RNA Folding Kinetics via a Ribosome Drafting Mechanism[J]. Journal of the American Chemical Society, 2016, 138(22):7016.

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