Composite
pBAD-GFP

Part:BBa_K845000:Design

Designed by: Grégory Hansen   Group: iGEM12_Grenoble   (2012-09-26)
Revision as of 15:40, 29 September 2012 by Greghansen (Talk | contribs) (→‎References)

iGEM 2012 Grenoble Team proposes a new design and application to the pBAD promoter(paraBAD).


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 500
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 344
    Illegal XhoI site found at 494
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 500
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 500
    Illegal AgeI site found at 179
    Illegal AgeI site found at 484
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1169
    Illegal SapI site found at 161


Design Notes

Design note: http://www.weizmann.ac.il/mcb/UriAlon/Papers/Zaslaver_Ecoli_library.pdf

Safety note: https://static.igem.org/mediawiki/2012/2/2a/ParaBAD-gfp.pdf

Source

The part comes from the plasmid collection Alon (http://www.weizmann.ac.il/mcb/UriAlon/Papers/Zaslaver_Ecoli_library.pdf). However, pBAD is a natural promoter in E. Coli. It regulates the production of three proteins (AraA, AraB and AraD) which form the arabinose operon. Their production enables the use of arabinose as a carbon source. It has six regulation sites, five of them are devoted to AraC fixation (three are repressing, one has a dual activity and one is an activator); the last one is a CRP binding site (positive activity). This promoter is activated when both activated CRP and AraC are bind to it.

GFP comes from the protein comes from Aequorea victoria. This jellyfish uses GFP (Green Fluorescent Protein) in order to convert the blue luminescence emitted by the aequorine into a green luminescence. Aequorea victoria: is a jellyfish that can be found off the coast of north America.

pBAD usage in biology: http://www.univ-orleans.fr/sciences/BIOCHIMIE/L/Illustrations%20cours/SLO-5BC03%20Regulation%20expression%20genome/Procaryotes/SLO-5BC03-COURS3.pdf

References

http://www.univ-orleans.fr/sciences/BIOCHIMIE/L/Illustrations%20cours/SLO-5BC03%20Regulation%20expression%20genome/Procaryotes/SLO-5BC03-COURS3.pdf

Zaslaver, A., Bren, A., Ronen, M., Itzkovitz, S., Kikoin, I., Shavit, S., Liebermeister, W., et al (2006). A comprehensive library of fluorescent transcriptional reporters for Escherichia coli. Nature Methods, 3(8), 623-628. doi:10.1038/nmeth895

http://2012.igem.org/Team:Grenoble/Biology/AND_gate