Coding

Part:BBa_K118016:Design

Designed by: Andrew Hall   Group: iGEM08_Edinburgh   (2008-10-06)
Revision as of 21:31, 6 October 2008 by Andhi (Talk | contribs) (→‎References)

glgC16 (glgC with G336D substitution)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 194
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The mutation G336D was achieved by the substitution of A for G at position 1076 (codon GGC->GAC) using the mutagenesis by blunt-ended ligation (MABEL) technique. The genomic DNA sequence also contains to EcoRI sites, both of which have been mutated out using MABEL.

Source

Escherichia coli JM109 genomic DNA

References

  • [http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=212844&blobtype=pdf Leung, P., Lee, Y. M., Greenberg, E., Esch, K., Boylan, S. and Preiss, J. (1986). Cloning and expression of the Escherichia coli glgC gene from a mutant containing an ADPglucose pyrophosphorylase with altered allosteric properties. J. Bacteriol. 167, 82-88.]
  • [http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WB5-45KKRWW-9P&_user=809099&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000043939&_version=1&_urlVersion=0&_userid=809099&md5=aa8d0cb9ae204115911c188cbdc31b16f Meyer, C. R., Bork, J. A., Nadler, S., Yirsa, J. and Preiss, J. (1998). Site-Directed Mutagenesis of a Regulatory Site of Escherichia coli ADP-Glucose Pyrophosphorylase: The Role of Residue 336 in Allosteric Behavior. Archives of Biochemistry and Biophysics 353, 152-159.]