Difference between revisions of "Part:BBa K3853010"
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<partinfo>BBa_K3853010 short</partinfo>
 
<partinfo>BBa_K3853010 short</partinfo>
  
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Hydrophobin-1 (HFB1) is a kind of class Ⅱ HFBs derived from <i>Trichoderma reesei</i>. 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<sup>[1]</sup>. 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_K3853054</partinfo> to construct the expression system to express and purify the protein.
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Hydrophobin-1 (HFB1) is a kind of class Ⅱ HFBs derived from <i>Trichoderma reesei</i>. This is the version with SpyTag. In order to constructed 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<sup>[1]</sup>. 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_K3853054</partinfo> to construct the expression system to express and purify the protein.
  
 
===Biology===
 
===Biology===

Revision as of 17:35, 17 October 2021


his-tag-SpyTag-HFB1

Hydrophobin-1 (HFB1) is a kind of class Ⅱ HFBs derived from Trichoderma reesei. This is the version with SpyTag. In order to constructed 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[1]. 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_K3853054 to construct the expression system to express and 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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]