Generator

Part:BBa_K2239008:Design

Designed by: Ruohong Wang   Group: iGEM17_SDSZ-China   (2017-10-01)


CBD-LDH


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1266
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 862
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1122


Design Notes

The CBD tag is added to purify the effective enzymes. When connected with the enzyme DNA and expressed, CBD is able to bind to the cellulose on gauze. After washing off other chemicals in the solution, the enzymes are left on the gauze. As a result, they are successfully immobilized.


Source

The LDH DNA is originally from the genome of Lactobacillus delbrueckii subsp. Bulgaricus, and is artificially synthesized based on the sequence retrieved from the GenBank.

The CBD sequence is retrieved from the GenBank and artificially synthesized.


References

[1] Ming-Min Zheng, Ru-Feng Wang, Chun-Xiu Li, Jian-He Xu: Two-step enzymatic synthesis of ursodeoxycholic acid with a new 7β-hydroxysteroid dehydrogenase from Ruminococcus torques. Process Biochemistry, Elsevier, 2015.

[2] Etai Shpigel, Arie Goldlust, Gilat Efroni, Amos Avraham, Adi Eshel, Mara Dekel, Oded Shoseyov: Immobilization of Recombinant Heparinase I Fused to Cellulose-Binding Domain, 1999.