Difference between revisions of "Part:BBa K3396000"

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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3396000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3396000 SequenceAndFeatures</partinfo>
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<p style=" font-weight: bold; font-size:20px;"> Improvement by CU_Egypt team 2022 </p>
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From literature we found that the yield of expression of Docs is very low so we decided to tag it by GST to increase its stability and the yield. Also, we optimized the sequence to be expressed in E-coli. in addition, there is no characterization for Docs on the registry so we expressed and characterized it by different methods such as Agarose gel electrophoresis, SDS PAGE, transformation efficiency, affinity chromatography, and Bradford assay.
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<p style=" font-weight: bold; font-size:14px;"> 1. Dry Lab </p>
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<p style=" font-weight: bold; font-size:12px;"> 1.1. Modeling </p>
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Docs has been tagged with GST and His for purification and increasing the yield by the GST tag. then the model designed by several tools to get the best model.
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<p style=" font-weight: bold; font-size:12px;"> Docs-GST </p>
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<html>
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<p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/doc-gst.png" style="margin-left:200px;" alt="" width="500" /></p>
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</html>
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                  Figure 1.: Predicted 3D structure of Docs-GST designed by RosettaFold tool.
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<html>
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<p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/doc-gst-score.png" style="margin-left:200px;" alt="" width="500" /></p>
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</html>
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                                Table 1.: QA scores of Docs-GST structure.
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===Reference===
 
===Reference===

Revision as of 20:03, 5 October 2022


DocS

The Coch2 module binds DocS (BBa_K3396000) modules constitutively.

Usage and Biology

The DocS[1] module comes from The C. thermocellum scaffoldin and it could recognize and bind tightly to complementary Coh2 modules. The Coh2–DocS pair represents the interaction between two complementary families of protein modules that exhibit divergent specificities and affinities, ranging from one of the highest known affinity constants between two proteins to relatively low-affinity interactions.


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]


Improvement by CU_Egypt team 2022


From literature we found that the yield of expression of Docs is very low so we decided to tag it by GST to increase its stability and the yield. Also, we optimized the sequence to be expressed in E-coli. in addition, there is no characterization for Docs on the registry so we expressed and characterized it by different methods such as Agarose gel electrophoresis, SDS PAGE, transformation efficiency, affinity chromatography, and Bradford assay.

1. Dry Lab

1.1. Modeling

Docs has been tagged with GST and His for purification and increasing the yield by the GST tag. then the model designed by several tools to get the best model.

Docs-GST 

                 Figure 1.: Predicted 3D structure of Docs-GST designed by RosettaFold tool.



                                Table 1.: QA scores of Docs-GST structure.




Reference

[1] BARAK Y, HANDELSMAN T, NAKAR D, et al. Matching fusion protein systems for affinity analysis of two interacting families of proteins: the cohesin-dockerin interaction [J]. J Mol Recognit, 2005, 18(6): 491-501.