Difference between revisions of "Part:BBa K2787013"
(3 intermediate revisions by 2 users not shown) | |||
Line 5: | Line 5: | ||
The 16S rRNA BBa_K2787040 under constitutive promoter BBa_J23100(BBa_K2787008) and pCons-oRBSB8-GFP(BBa_K2787001) are linked into one plasmid to quantify the orthogonality of the TJU and B8 ribosome system. | The 16S rRNA BBa_K2787040 under constitutive promoter BBa_J23100(BBa_K2787008) and pCons-oRBSB8-GFP(BBa_K2787001) are linked into one plasmid to quantify the orthogonality of the TJU and B8 ribosome system. | ||
− | |||
===Usage and Biology=== | ===Usage and Biology=== | ||
− | + | The plasmid with GFP under C9 orthogonal RBS with orthogonal 16S rRNA A2 is constructed. | |
+ | [[File:Shanghaitech2018-oribo-orbs-07-07.png|700px|center]] | ||
+ | [[File:ShanghaiTech2018-b8 rbs.png|700px|center]] | ||
+ | The GFP under control of B8 orthogonal RBS is tested within different 16s rRNAs. The fluorescence with C9 orthogonal ribosome is higher than host ribosomes and A2,TJU ribosomes, which means the orthogonality between that of C9 ribosome is not as expected. | ||
+ | [[File:ShanghaiTech-TJU_16S.png|700px|center]] | ||
+ | The GFP under control of different orthogonal RBS is tested within TJU orthogonal 16s rRNAs. There is no fluorescence difference between each group, means that the A2 ribosomes is orthometric to other RBSs. | ||
+ | [[File:A2-B8-C9-Orthogonality.png|700px|center]] | ||
+ | The expression of the gene of interest is tested between different orthogonal RBS and different orthogonal ribosomes. | ||
<!-- --> | <!-- --> | ||
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> |
Latest revision as of 03:43, 18 October 2018
GFP under B8 orthogonal RBS with orthogonal 16S rRNA BBa_K2787041
The 16S rRNA BBa_K2787040 under constitutive promoter BBa_J23100(BBa_K2787008) and pCons-oRBSB8-GFP(BBa_K2787001) are linked into one plasmid to quantify the orthogonality of the TJU and B8 ribosome system.
Usage and Biology
The plasmid with GFP under C9 orthogonal RBS with orthogonal 16S rRNA A2 is constructed.
The GFP under control of B8 orthogonal RBS is tested within different 16s rRNAs. The fluorescence with C9 orthogonal ribosome is higher than host ribosomes and A2,TJU ribosomes, which means the orthogonality between that of C9 ribosome is not as expected.
The GFP under control of different orthogonal RBS is tested within TJU orthogonal 16s rRNAs. There is no fluorescence difference between each group, means that the A2 ribosomes is orthometric to other RBSs.
The expression of the gene of interest is tested between different orthogonal RBS and different orthogonal ribosomes. 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 798
Illegal NheI site found at 821 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1661
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1675
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 707