Composite

Part:BBa_K2027001:Design

Designed by: Michael Becich   Group: iGEM16_Stanford-Brown   (2016-09-07)
Revision as of 06:13, 24 October 2016 by Theresasievert (Talk | contribs)


AE Blue-CBD-FlagLumioHisTag Fusion Protein

aeBlue-CBD-His-tag is a composite part created using the aeBlue chromoprotein from the iGEM registry BBa_K1033929 and adding a cellulose binding domain, and FLAG, lumino and 6x histidine tag. The aeBlue chromoprotein has been extracted from the sea anemone, Actinia equine [1]. The cellulose binding domain (CBD) is from the BioBrick BBa_K1321366 which also includes a GFP fused to it. For the purposes of our experiments, we isolated the cellulose binding domain from this part using PCR. The cellulose binding domain sequence was isolated from Cellulomonas fimi and has been shown to bind irreversibly to cellulose [2]. The FLAG, lumino and 6x histidine tag was synthesized for use in this part. This part is able to bind to cellulose and also be purified using the FLAG, lumino and 6x histidine tag.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 740


References

1. Shkrob, Maria A., Yurii G. Yanushevich, Dmitriy M. Chudakov, Nadya G. Gurskaya, Yulii A. Labas, Sergey Y. Poponov, Nikolay N. Mudrik, Sergey Lukyanov, and Konstantin A. Lukyanov. "Far-red Fluorescent Proteins Evolved from a Blue Chromoprotein from Actinia Equina." Biochem. J. Biochemical Journal 392.3 (2005): 649-54. Web. 2. Carrard, G., A. Koivula, H. Soderlund, and P. Beguin. "Cellulose-binding Domains Promote Hydrolysis of Different Sites on Crystalline Cellulose." Proceedings of the National Academy of Sciences 97.19 (2000): 10342-0347. Web.