Team:INSA-Lyon/Project/Stage2/Results
From 2010.igem.org
(Difference between revisions)
Line 44: | Line 44: | ||
<h2>Results</h2> | <h2>Results</h2> | ||
<p> | <p> | ||
- | <br><h3><font color="purple">Observation of the phasin-intein fusion protein (BBa_K342002)on the granule</font></h3><br><br> | + | <br><h3><font color="purple">Observation of the phasin-intein fusion protein (BBa_K342002) on the granule</font></h3><br><br> |
<p>Our ligation didn't work out so we couldn't realize the observations as planned.<br/> | <p>Our ligation didn't work out so we couldn't realize the observations as planned.<br/> | ||
We had many difficulties to add the constitutive promoter despite numerous tries.</p> <br><br> | We had many difficulties to add the constitutive promoter despite numerous tries.</p> <br><br> | ||
Line 50: | Line 50: | ||
<p>We made a double transformation with pILI1 and 20N: The resultant bacteria synthesize the PHB drops and over synthesize the licopen. | <p>We made a double transformation with pILI1 and 20N: The resultant bacteria synthesize the PHB drops and over synthesize the licopen. | ||
<br/><br/> | <br/><br/> | ||
- | <p>When we | + | <p>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.<br/> |
<ol id="list_extra"> | <ol id="list_extra"> | ||
<li>Serie 1 : in LB media with glucose 7% and 2,5µg of RedNile<br/> | <li>Serie 1 : in LB media with glucose 7% and 2,5µg of RedNile<br/> | ||
<ol id="list_extra"> | <ol id="list_extra"> | ||
<li> pILI1 with ampiciline</li> | <li> pILI1 with ampiciline</li> | ||
- | <li>double transformation with | + | <li>double transformation with ampicillin</li> |
- | <li>double transformation with | + | <li>double transformation with ampicillin and kanamycin</li> |
- | <li>The lipid part with | + | <li>The lipid part with kanamycin</li> |
</ol><br> | </ol><br> | ||
<li>Serie 2 : in LB media with 2,5µg of RedNile<br/> | <li>Serie 2 : in LB media with 2,5µg of RedNile<br/> | ||
<ol id="list_extra"> | <ol id="list_extra"> | ||
- | <li> pILI1 with | + | <li> pILI1 with ampicillin</li> |
- | <li>double transformation with | + | <li>double transformation with ampicillin</li> |
- | <li>double transformation with | + | <li>double transformation with ampicillin and kanamycin</li> |
- | <li>The lipid part with | + | <li>The lipid part with kanamycin</li> |
</ol><br> | </ol><br> | ||
<li>Serie 3 : in LB media with glucose 7%<br/> | <li>Serie 3 : in LB media with glucose 7%<br/> | ||
<ol id="list_extra"> | <ol id="list_extra"> | ||
- | <li> pILI1 with | + | <li> pILI1 with ampicillin</li> |
- | <li>double transformation with | + | <li>double transformation with ampicillin and kanamycin</li> |
- | <li>The lipid part with | + | <li>The lipid part with kanamycin</li> |
</ol><br> | </ol><br> | ||
- | <li>Serie 4 : in LB media, the double transformation with | + | <li>Serie 4 : in LB media, the double transformation with ampicillin and kanamycin</li> |
</ol><br/> | </ol><br/> | ||
Latest revision as of 00:36, 28 October 2010
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.
- Serie 1 : in LB media with glucose 7% and 2,5µg of RedNile
- pILI1 with ampiciline
- double transformation with ampicillin
- double transformation with ampicillin and kanamycin
- The lipid part with kanamycin
- Serie 2 : in LB media with 2,5µg of RedNile
- pILI1 with ampicillin
- double transformation with ampicillin
- double transformation with ampicillin and kanamycin
- The lipid part with kanamycin
- Serie 3 : in LB media with glucose 7%
- pILI1 with ampicillin
- double transformation with ampicillin and kanamycin
- The lipid part with kanamycin
- 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.