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Part:BBa_K1355002

Designed by: Maria Clara Astolfi, Luna Barroco de Lacerda   Group: iGEM14_UFAM_Brazil   (2014-10-06)
Revision as of 03:48, 17 October 2014 by Lunalacerda (Talk | contribs)

Mercury ions detector device

We designed a biobrick device to express Green Fluorescent Protein in mercury’s occurrence. The Mercury ions’ detector device biobrick (BBa_K1355002) is composed by mer bidirectional promoter (BBa_K1355001) attached to the GFP translational unit (BBa_E0840) available on iGEM database since 2004 . It has dual function: A) In reverse: MerR protein regulator transcription; and B) in forward: transcription of MerP - MerT - GFP proteins, as represented below:


L4.jpg

In absence of mercury, MerR forms a MerR-promoter-operator complex, preventing RNA polymerase to recognize the promoter, consequently, messengers RNA for MerPT and GFP will not be transcript. In presence of Hg2+, MerR protein binds to this element and dissociates from the promoter-operator complex, allowing MerPT and GFP expression, as represented below:

Bs4.png

When MerT, MerP and the Green Fluorescent Protein (GFP) protein are expressed, the biodetection will start! MerP and MerT proteins are responsible for the transport of mercury from the periplasm to cytoplasm and GFP will act as a reporter alerting the presence of mercury in the middle and higher is mercury’s concentration, higher will be GFP expression.


Biosensor.jpg

Figure 1: Mercury Bacter Hg biodetector (DH5-alpha transformed with BBa_K1355002)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 988
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 586
    Illegal NgoMIV site found at 1160
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1862
    Illegal SapI site found at 579


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