Difference between revisions of "Part:BBa K3853009"
Lp-tiffany (Talk | contribs) |
Lp-tiffany (Talk | contribs) |
||
| Line 5: | Line 5: | ||
Aryl alcohol oxidase (AAO) is an enzyme containing flavin-adenine-dinucleotide (FAD). This is the version with SpyTag. In order to be able to construct our multi-enzyme complex, we introduced the SpyTag/SpyCatcher system. We added SpyTag to the N-terminus of each protein through 10 × ELP, which is a oligopeptide linker that does not affect the function of the protein it attaches to, as described in the literature. So this part can be combined with dCas9-SpyCatcher through covalent isopeptide action, thereby being immobilized on dsDNA. His-tag was added to purify the protein. We use <partinfo>BBa_K3853053</partinfo> to construct the expression system and to express and to purify the protein. | Aryl alcohol oxidase (AAO) is an enzyme containing flavin-adenine-dinucleotide (FAD). This is the version with SpyTag. In order to be able to construct our multi-enzyme complex, we introduced the SpyTag/SpyCatcher system. We added SpyTag to the N-terminus of each protein through 10 × ELP, which is a oligopeptide linker that does not affect the function of the protein it attaches to, as described in the literature. So this part can be combined with dCas9-SpyCatcher through covalent isopeptide action, thereby being immobilized on dsDNA. His-tag was added to purify the protein. We use <partinfo>BBa_K3853053</partinfo> to construct the expression system and to express and to purify the protein. | ||
| + | ===Biology=== | ||
| − | + | ===Usage=== | |
| − | ===Usage | + | |
| + | ===References=== | ||
| + | [1] Lim, S., Kim, J., Kim, Y., Xu, D. & Clark, D. S. CRISPR/Cas-directed programmable assembly of multi-enzyme complexes. <i>Chemical communications (Cambridge, England)</i> <b>56</b>, 4950-4953, doi:10.1039/d0cc01174f (2020). | ||
<!-- --> | <!-- --> | ||
Revision as of 18:53, 15 October 2021
his-tag-SpyTag-AAO
Aryl alcohol oxidase (AAO) is an enzyme containing flavin-adenine-dinucleotide (FAD). This is the version with SpyTag. In order to be able to construct our multi-enzyme complex, we introduced the SpyTag/SpyCatcher system. We added SpyTag to the N-terminus of each protein through 10 × ELP, which is a oligopeptide linker that does not affect the function of the protein it attaches to, as described in the literature. So this part can be combined with dCas9-SpyCatcher through covalent isopeptide action, thereby being immobilized on dsDNA. His-tag was added to purify the protein. We use BBa_K3853053 to construct the expression system and to express and to purify the protein.
Biology
Usage
References
[1] Lim, S., Kim, J., Kim, Y., Xu, D. & Clark, D. S. CRISPR/Cas-directed programmable assembly of multi-enzyme complexes. Chemical communications (Cambridge, England) 56, 4950-4953, doi:10.1039/d0cc01174f (2020).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 699
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 412
Illegal BamHI site found at 1411
Illegal BamHI site found at 1798 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]