Difference between revisions of "Part:BBa K1355002"

Revision as of 03:48, 17 October 2014

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:

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:

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.

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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 <partinfo>BBa_K1355002 short</partinfo>
+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:
-We used the biobrick (BBa_E0840) available on iGEM database since 2004 to connect to our “essential bio brick” (BBa_K1355001) and - together - form a system capable of bio detection mercury in bacteria, represented below:
 [[File:L4.jpg]]
-In the presence of mercury, MerR protein will bind to it and MerT, MerP and the Green Fluorescent Protein (GFP) protein will be expressed and bio detection starts! 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. And if there isn’t mercury on medium, MerT, MerP and GFP synthesis will be repressed by MerR repressor protein.
+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:
+[[File: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.
 [[File:biosensor.jpg]]