Coding
Biosurfact

Part:BBa_K653000:Design

Designed by: Team Panama 2011   Group: iGEM11_Panama   (2011-09-26)
Revision as of 03:12, 27 September 2011 by Ernestopro (Talk | contribs) (→‎References)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

Re-designing "The Biosurfactator" The first Central American BioBrick


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 103
    Illegal BamHI site found at 663
    Illegal XhoI site found at 839
    Illegal XhoI site found at 2125
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1018
    Illegal NgoMIV site found at 1739
    Illegal NgoMIV site found at 1852
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 328
    Illegal BsaI site found at 1368
    Illegal BsaI.rc site found at 512


Design Notes

The RhlAB gene complex has shown illegal restriction sites according to the assembly standard protocol 10. We faced this problem by applying the mutagenesis protocol that consist in doing silent mutations that change the nucleotides of our rhlAB gene that are cut by the Pstl restriction enzyme. This will give us a standardized coding sequence of the rhlAB gene compatible with the Assembly Standard protocol 10 and a new BioBrick part for the Registry.



Source

Pseudomonas aeruginosa from INDICASAT lab. and the genomic sequence of RhlAB was reviwed in genBank (ID 878955 for rhamnosyltransferase A and ID 878954 for rhamnosyltransferase B)


http://www.ncbi.nlm.nih.gov/gene?term=RhlA%20ID%20878955

http://www.ncbi.nlm.nih.gov/gene?term=RhlB%20ID%20878954

http://www.indicasat.org.pa/

References

Qinhong Wang, Xiangdong Fang, Baojun Bai, Xiaolin Liang, Patrick J. Shuler, William A. Goddard III, Yongchun Tang. 2007. Engineering Bacteria for Production of Rhamnolipid as an Agent for Enhanced Oil Recovery. Biotechnology and Bioengineering Volume 98, Issue 4, pages 842–853.

Ochsner UA, Fiechter A, Reiser J. 1994. Isolation, characterization, and expression in Escherichia coli of the Pseudomonas aeruginosa rhlAB genes encoding a rhamnosyltransferase involved in rhamnolipid biosurfactant synthesis. J Biol Chem 269:19787–19795.

Maier RM, Soberon-Chavez G. 2000. Pseudomonas aeruginosa rhamnolipids: biosynthesis and potential applications. Appl Microbiol Biotechnol 54(5):625–633.

QuikChange Lightning Multi Site-Directed Mutagenesis Kit Instruction manual, catalog #210515 from Stratagene.