Part:BBa_K2787009
GFP under A2 orthogonal RBS with orthogonal 16S rRNA A2
The 16S rRNA A2 under constitutive promoter BBa_J23100(BBa_K2787006) and pCons-oRBSA2-GFP(BBa_K2787000) are linked into one plasmid to justify the proper function of the ribosomal 16SRNA and RBS set A2.
Usage and Biology
a plasmid with GFP under control of A2 orthogonal RBS with A2 orthogonal 16s rRNA is constructed to test the orthogonality of A2 RBS.
GFP, the target gene downstream the A2 orthogonal RBS is highly activated by the A2 orthogonal ribosome. The expression of GFP increases gradually over time. In the absence of A2 orthogonal ribosome, the GFP is repressed deeply, which shows the high orthogonality of A2 orthogonal RBS and A2 orthogonal ribosome.
The GFP under control of A2 orthogonal RBS is tested within different 16s rRNAs. The fluorescence with A2 orthogonal ribosome is the best among host ribosomes and A2,B8,C9 ribosomes, which shows the good orthogonality of A2 16s rRNA with A2 orthogonal RBS.
The fluorescence under the test with 3 different orthogonal RBS and 3 different orthogonal ribosmes. It shows that the orthogonality of A2 ribosome and A2 RBS is one of the best ones.
[[File:Shanghaitech-A2.png|450px|centerFig. 3
A2 O-16S fluorescence at 200 minutes, showing that the exist of A2 orthogonal 16s rRNA will greatly activate the GFP. However, it will be deeply depressed without the A2 ribosome.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 797
Illegal NheI site found at 820 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1659
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 706
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