Generator

Part:BBa_K2239001:Design

Designed by: Ruohong Wang   Group: iGEM17_SDSZ-China   (2017-09-30)
Revision as of 14:20, 31 October 2017 by ChrisWang (Talk | contribs) (→‎References)

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ChBD-7alphaHSDH


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1041
  • 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 875


Design Notes

The ChBD tag is added to purify the effective enzymes. When connected with the enzyme DNA and expressed, ChBD is able to bind to the chitin powder. As a result, the enzyme is successfully immobilized on the chitin powder inside the solution.


Source

The 7alpha-HSDH sequence is cloned from the genome of E.coli DH5alpha through PCR amplification, using the primers designed and synthesized based on its sequence.

The ChBD 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] Takahisa Ikegami, Terumasa Okada, Masayuki Hashimoto, Shizuka Seino, Takeshi Watanabe, and Masahiro Shirakawa: Solution Structure of the Chitin-binding Domain of Bacillus circulans WL-12 Chitinase A1. The Journal Of Biological Chemistry, 2000.