Part:BBa_K2540015
Orthogonal RBS for expression across bacteria
This orthogonal RBS was designed using RBS calculator for orthogonal 16S rRNA predicted to function across a variety of bacterial strains. Prediction was done by algorithm written by Rice iGEM 2018 team based on previous work by Chubiz & Rao.
Usage and Biology
The orthogonal RBS contains an altered Shine-Dalgarno sequence, which prevents binding of the wild-type 16S rRNA subunit. Only 16S rRNA subunits containing a complementary orthogonal anti-Shine Dalgarno sequence may bind to the orthogonal RBS. This RBS along with its corresponding 16S rRNA may be used when orthogonal translation is desired in bacteria.
Figure 11: characterization of orthogonal translation constructs in E. coli . IPTG-dependent mKate2 fluorescence is observed when plamids containing o16S rRNA controlled by Plac promoter and oRBS-mKate2 are co-transformed into E. coli (left). At 0.1 mM IPTG, ~100 fold fluorescence over negative control is observed, demonstrating that translation is orthogonal (right).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3
Illegal BsaI.rc site found at 58
References
Chubiz, L. M., & Rao, C. V. (2008). Computational design of orthogonal ribosomes. Nucleic Acids Research, 36(12), 4038–4046.
| None |