Difference between revisions of "Part:BBa K4814013"
 
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To optimize the design of the RecA-eGFP reporter (BBa_K3020000), we replaced the original B0034 RBS with a new RBS designed by Zhang, M. et al (2022), and see if the effectiveness will increase. However, from our experimental result, using RBS BBa_K4814012 does not enhance the level of fluorescence of the reporter.
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To improve the RecA (K3) - B0034 - EGFP bioreporter we designed to test RecA(K3) (BBa_K3020001) promoter, we replaced the original B0034 RBS with a new RBS designed by Zhang, M. et al (2022), and see if the effectiveness will increase. However, from our experimental result, using RBS BBa_K4814012 (Strong) does not enhance the level of fluorescence of the reporter.
  
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We also tried different RBS (strong and B0032) to replace the B0034 (BBa_K4814015), and the result indicated that RecA(K3)-eGFP with B0032 RBS has a stronger SFU (FL over OD600) than the original B0034 we chose. Please refer to BBa_K4814002 (B0032) and BBa_K4814013 (Strong RBS) for details.
 
 
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Latest revision as of 09:55, 12 October 2023


RecA(K3)-Strong-eGFP

To improve the RecA (K3) - B0034 - EGFP bioreporter we designed to test RecA(K3) (BBa_K3020001) promoter, we replaced the original B0034 RBS with a new RBS designed by Zhang, M. et al (2022), and see if the effectiveness will increase. However, from our experimental result, using RBS BBa_K4814012 (Strong) does not enhance the level of fluorescence of the reporter.

RBS test of UVB and H2O2 FL over OD600 of RecA(K3)-B0034-eGFP, RecA(K3)-B0032-eGFP, and RecA(K3)-Strong-eGFP.

RBS test of Nalidixic acid and Aspartame FL over OD600 of RecA(K3)-B0034-eGFP, RecA(K3)-B0032-eGFP, and RecA(K3)-Strong-eGFP.


Zhang, M., Holowko, M. B., Zumpe, H. H., & Ong, C. S. (2022). Machine Learning Guided Batched Design of a Bacterial Ribosome Binding Site. ACS Synthetic Biology, 11(7), 2314-2326. https://doi.org/10.1021/acssynbio.2c00015

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 NgoMIV site found at 847
  • 1000
    COMPATIBLE WITH RFC[1000]