Regulatory

Part:BBa_K1935008

Designed by: Jean DESCARPENTRIE   Group: iGEM16_Bordeaux   (2016-10-13)


pMYO-2 : Specific Pharynx muscle promoter of C.elegans

myo-2 gene promoter of Caenorhabditis elegans basically drives the expression of MYO-2 in pharynx muscle of C.elegans.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 580
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 490


Usage and Biology

In Caenorhabditis elegans, we can do the transformation by micro-injection in order to create mutant worm. However, even if we microinjected a worm, we cannot be sure that the microinjection and the transformation succeeded. This is why we need some benchmark to identify transformed worms and non transformed worms. This bechmark can be pMyo2::GFP. It will allow the researcher to screen the transformed worms thanks to their green fluorescent pharynx muscle. For that, a mix has to be prepared. In this mix,a plasmid control is added to the plasmids of the study. Myo2 with GFP can be used for that. Indeed the fluorescence of the GFP can be observed for control the worms correctly injected.

Fluorescent pharynx muscle in C.elegans micro-injected with pMyo2::GFP

C.elegans eats the bacteria and fungi. When there are no more bacteria on the petri dish, the worms are starved. It can be useful to see if the starvation can change a phenotype, as the worm can be adaptive.

In order to characterize our promoter pMyo2, we compared the fluorescence in the pharynx between starved worms and non starved worms.

Fluorescent pharynx muscle in C.elegans micro-injected with pMyo2::GFP

We calculated the corrected total pharynx fluorescence of 10 worms from each sample, starved and not starved. We calculated then the mean of this florescence. As a result, we observe a higher fluorescence in the feed sample.

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