Difference between revisions of "Part:BBa K2052015:Design"

(→‎References)
(→‎Design Notes)
 
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===Design Notes===
 
===Design Notes===
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RBS and our protein coding region (ButCoaT) are digested from our whole construct with the enzymes Sall and HindIII.
+
RBS and our protein coding region (ButCoaT) are digested from our whole construct with the enzymes Xbal and PstI.
  
  
−
In order to be expressed in E.coli BL21, the sequence was optimized with the use of codon bias rule and the unwanted restriction cut sides(SalI, HindIII) were removed with point mutation.
+
In order to be expressed in E.coli BL21, the sequence was optimized with the use of codon bias rule and the unwanted restriction cut sides(XbaI, PstI) were removed with point mutation.
  
 
===Source===
 
===Source===

Latest revision as of 01:15, 20 October 2016


ButCoat with strong RBS


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]


Design Notes

RBS and our protein coding region (ButCoaT) are digested from our whole construct with the enzymes Xbal and PstI.


In order to be expressed in E.coli BL21, the sequence was optimized with the use of codon bias rule and the unwanted restriction cut sides(XbaI, PstI) were removed with point mutation.

Source

Source of ButCoaT Gene: Roseburia intestinalis L1-82

References

Hassig, C.A., Tong, J.K., Schreiber, S.L. (1997). Fiber-derived Butyrate and the Prevention of Colon Cancer Roberto Berni Canani, Margherita Di Costanzo, Ludovica Leone, Monica Pedata, Rosaria Meli, Antonio Calignano. (2011). Potential beneficial affects of butyrate in intestinal and extraintestinal diseases Duncan,S. H., Barcenilla, A., Stewart, C. S., Pryde, S. E., Flint, H. J. (2002). Acetate Utilization and Butyryl Coenzyme A (CoA):Acetate-CoA Transferase in ButyrateProducing Bacteria from the Human Large Intestine Sequence : http://www.uniprot.org/uniprot/C7GB37?sort=score Pathway : http://www.genome.jp/dbget-bin/www_bget?reaction+R00393+R01179+R01359+R07832 Picture : http://www.brenda-enzymes.org/structure.php?show=reaction&id=360609&type=S&displayType=marvin