Team:INSA-Lyon/Project/Stage2/Results

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Results


Observation of the phasin-intein fusion protein (BBa_K342002) on the granule



Our ligation didn't work out so we couldn't realize the observations as planned.
We had many difficulties to add the constitutive promoter despite numerous tries.




Lipidic inclusion in the PHB drop



We made a double transformation with pILI1 and 20N: The resultant bacteria synthesize the PHB drops and over synthesize the licopen.

When we obtained the clones, we started four different series of cultures in order to analyze the will of the lipid when the granule are produced.

  1. Serie 1 : in LB media with glucose 7% and 2,5µg of RedNile
    1. pILI1 with ampiciline
    2. double transformation with ampicillin
    3. double transformation with ampicillin and kanamycin
    4. The lipid part with kanamycin

  2. Serie 2 : in LB media with 2,5µg of RedNile
    1. pILI1 with ampicillin
    2. double transformation with ampicillin
    3. double transformation with ampicillin and kanamycin
    4. The lipid part with kanamycin

  3. Serie 3 : in LB media with glucose 7%
    1. pILI1 with ampicillin
    2. double transformation with ampicillin and kanamycin
    3. The lipid part with kanamycin

  4. Serie 4 : in LB media, the double transformation with ampicillin and kanamycin

We observed a sample of each culture under fluorescence microscope. Unfortunately we couldn't see the difference between the granules and the licopen.


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