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Team:SDU-Denmark/Tour42
From 2014.igem.org
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| - | The pTet expression system and limonene synthase construct is evolved around one thing: the OneProt. | + | <span class="intro">The pTet expression system</span> and limonene synthase construct is evolved around one thing: the OneProt. |
We have made the pTet-OneProt construct in order for us to synthesize a nutritional protein with the | We have made the pTet-OneProt construct in order for us to synthesize a nutritional protein with the | ||
correct ratio of essential amino acids and the correct ratio between essential and non-essential | correct ratio of essential amino acids and the correct ratio between essential and non-essential | ||
amino acids. The device is found as <a href="http://parts.igem.org/Part:BBa_K1475000"> Bba_K1475000.</a><br><br> | amino acids. The device is found as <a href="http://parts.igem.org/Part:BBa_K1475000"> Bba_K1475000.</a><br><br> | ||
| - | The protein is self-designed, so we wanted to test if the protein were expressed in <i>E. coli</i> K12 MG1655, by | + | <span class="intro">The protein is self-designed,</span> so we wanted to test if the protein were expressed in <i>E. coli</i> K12 MG1655, by |
the use of Western blotting. The western blot was blottet with <i>E. coli</i> K12 MG1655 wild-type and <i>E. coli</i> | the use of Western blotting. The western blot was blottet with <i>E. coli</i> K12 MG1655 wild-type and <i>E. coli</i> | ||
expressing OneProt at different OD measures. | expressing OneProt at different OD measures. | ||
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| - | The protein has a 3xFLAG tag and since bonds are showing, OneProt is expressed. However, from this | + | <span class="intro">The protein has a</span> 3xFLAG tag and since bonds are showing, OneProt is expressed. However, from this |
western blot, we cannot see if the protein has been cut, just that it is expressed. | western blot, we cannot see if the protein has been cut, just that it is expressed. | ||
<br><br><br> | <br><br><br> | ||
Revision as of 03:04, 17 October 2014
OneProt
The pTet expression system and limonene synthase construct is evolved around one thing: the OneProt.
We have made the pTet-OneProt construct in order for us to synthesize a nutritional protein with the
correct ratio of essential amino acids and the correct ratio between essential and non-essential
amino acids. The device is found as Bba_K1475000.
The protein is self-designed, so we wanted to test if the protein were expressed in E. coli K12 MG1655, by
the use of Western blotting. The western blot was blottet with E. coli K12 MG1655 wild-type and E. coli
expressing OneProt at different OD measures.
Western blot showing E. coli wild-type and E. coli expressing OneProt at OD at 0.3, 0.8, 1.8 and an overnight
culture. The membrane has been exposed for 10 minutes.
The protein has a 3xFLAG tag and since bonds are showing, OneProt is expressed. However, from this
western blot, we cannot see if the protein has been cut, just that it is expressed.
