Difference between revisions of "Part:BBa K2239001:Design"

 
Line 7: Line 7:
  
 
===Design Notes===
 
===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.
  
  
Line 13: Line 13:
 
===Source===
 
===Source===
  
−
The 7alpha-HSDH sequence is cloned from the genome of E.coli through PCR amplification, using the primers designed and synthesized based on its sequence. The restriction site BamH I is added to the upstream primer, and Hind III is added on the downstream primer.
+
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 CBD sequence is retrieved from the GeneBank, and is inserted into plasmid pUC57. The CBD gene is then cloned from the plasmid by PCR, with the restriction site Hind III added to the upstream primer, and Xhol I added to the downstream primer.
+
 
 +
The ChBD sequence is retrieved from the GenBank and artificially synthesized.
 +
 
  
 
===References===
 
===References===

Revision as of 14:19, 31 October 2017


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