Difference between revisions of "Part:BBa K2623021"
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| − | As mentioned above, SpyTag was constructed in order to anchore L7Ae to outer membrane through the bioconjugation of the SpyTag/SpyCatcher system. We also construct another form of BBa_K2623021-<partinfo>BBa_K2623023</partinfo> The only difference is that we inserted SpyTag to C-termini of OmpA protein(instead of the N-termi in BBa_K2623021). Through the fluorescence intensity of GFP, we can compare and evaluate expression efficiency of SpyTag between the BBa_K2623021 and this part. Then the better one would be uesd to construct our final gene circuit. | + | As mentioned above, SpyTag was constructed in order to anchore L7Ae to outer membrane through the bioconjugation of the SpyTag/SpyCatcher system. We also construct another form of BBa_K2623021-<partinfo>BBa_K2623023</partinfo>. The only difference is that we inserted SpyTag to C-termini of OmpA protein(instead of the N-termi in BBa_K2623021). Through the fluorescence intensity of GFP, we can compare and evaluate expression efficiency of SpyTag between the BBa_K2623021 and this part. Then the better one would be uesd to construct our final gene circuit. |
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Revision as of 07:45, 13 October 2018
Bacterial outer membrane protein A (OmpA) fused with SpyTag at its N-termini and GFP at its C-termin
Summary
This part is a relatively basic part designed to target the archaeal ribosomal protein protein L7Ae and then siRNA into outer-membrane vesicles (OMVs). Porin outer-membrane protein A, OmpA, is a kind of highly expressed transmembrane porin protein of E. coli and is therefore abundant in OMVs. We inserted SpyTag to its N-termini to bioconjugate with the SpyCather in the following parts. In order to test the efficiency of anchoring the OmpA-SpyTag (OmpA-ST) in our circuit, we use GFP, BBa_I13401 as the reporter, because the fluorescence intensity is easier to be measured.
Usage and Biology
As mentioned above, SpyTag was constructed in order to anchore L7Ae to outer membrane through the bioconjugation of the SpyTag/SpyCatcher system. We also construct another form of BBa_K2623021-BBa_K2623023. The only difference is that we inserted SpyTag to C-termini of OmpA protein(instead of the N-termi in BBa_K2623021). Through the fluorescence intensity of GFP, we can compare and evaluate expression efficiency of SpyTag between the BBa_K2623021 and this part. Then the better one would be uesd to construct our final gene circuit.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 65
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1302