Difference between revisions of "Part:BBa K2123201"
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With this part, you can switch: I) promoter sequences, regulating (or not) by what you want; II) transporters protein, using the preferential one for your chassis; III) and whatever you want related to mercury metabolism! These two enzymes together increase the mercury metabolism spectrum in bacteria, improving bioremediation process. Check how the enzymatic pathway works on “Structure and mechanism” and it’s results in “Usage, Methodology and Experiments”.  
 
With this part, you can switch: I) promoter sequences, regulating (or not) by what you want; II) transporters protein, using the preferential one for your chassis; III) and whatever you want related to mercury metabolism! These two enzymes together increase the mercury metabolism spectrum in bacteria, improving bioremediation process. Check how the enzymatic pathway works on “Structure and mechanism” and it’s results in “Usage, Methodology and Experiments”.  
  
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==Structure and mechanism: ==
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==Usage, Methodology and Experiments==
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The first step to characterize this part was testing its Hg resistance and bioremediation with and without MerB gene, as represented below, through an inhibition zone.
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<center>https://static.igem.org/mediawiki/parts/c/cf/UFAM_MERBA_2.png</center>
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It has been use a 10 times concentration variation (20mg/mL, 200µg/mL and 20µg/mL) of HgCl2 in LM (Luria-Bertani variation with half salt) solid media, adding 10µL of mercury chloride solution on its paper disks. The samples were inoculated in triplicate and incubate in BOD at 37°C for 2 days. The results are shown below.
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<center>https://static.igem.org/mediawiki/parts/6/68/UFAM_MERBA_3.png</center>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 19:52, 27 October 2016


Strong RBS + MerB (Organomercurial Lyase) + Strong RBS + MerA (Mercuric Reductase) + B0015

Overview

This composite part was developed to turn available two mer operon enzymes, related to Hg metabolism: organomercurial lyase (MerB) and mercury reductase (MerA), as you can see below.

UFAM_MERBA_1.png

With this part, you can switch: I) promoter sequences, regulating (or not) by what you want; II) transporters protein, using the preferential one for your chassis; III) and whatever you want related to mercury metabolism! These two enzymes together increase the mercury metabolism spectrum in bacteria, improving bioremediation process. Check how the enzymatic pathway works on “Structure and mechanism” and it’s results in “Usage, Methodology and Experiments”.

Structure and mechanism:

Usage, Methodology and Experiments

The first step to characterize this part was testing its Hg resistance and bioremediation with and without MerB gene, as represented below, through an inhibition zone.

UFAM_MERBA_2.png

It has been use a 10 times concentration variation (20mg/mL, 200µg/mL and 20µg/mL) of HgCl2 in LM (Luria-Bertani variation with half salt) solid media, adding 10µL of mercury chloride solution on its paper disks. The samples were inoculated in triplicate and incubate in BOD at 37°C for 2 days. The results are shown below.

UFAM_MERBA_3.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 458
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 56
    Illegal NgoMIV site found at 630
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 49