"
Team:Trieste/notebook2
From 2012.igem.org
(Difference between revisions)
Corsogiulia (Talk | contribs) |
|||
| Line 11: | Line 11: | ||
<div id="suicide" class="notebook_section"> | <div id="suicide" class="notebook_section"> | ||
<h2 class="notebook_title">Suicide System</h2> | <h2 class="notebook_title">Suicide System</h2> | ||
| - | Finally we got the antimicrobial peptide LL37 in pEX-A plasmid and we amplified it through the transformation of DH5-alpha bacteria.</br> | + | Finally we got by synthesis the antimicrobial peptide LL37 cathelicidin (BBa_K875009) in pEX-A plasmid and we amplified it through the transformation of DH5-alpha bacteria.</br> |
</br> | </br> | ||
| - | The positive colonies were then digested EcoRI/SpeI, the fragment was eluted from the gel and then cut it again with EcoRI, because the pEX-A plasmid | + | The positive colonies were then digested EcoRI/SpeI, the fragment was eluted from the gel and then cut it again with EcoRI, because the pEX-A plasmid has another EcoRI restriction site downstream our fragment.</br> |
</br> | </br> | ||
| - | Then we did a column purification of the fragment and we successfully cloned the LL | + | Then we did a column purification of the fragment and we successfully cloned the LL 37 into the pSB1C3 linearized backbone to make it ready for a potential submission.</br> |
</br> | </br> | ||
| - | We cut | + | We LL 37 -previously cut EcoRI/SpeI- was cloned upstream the TT_B0015 and the DH5alpha cells were transformed with the plasmid (LL-37-B0015). The positive colonies were inoculated and then cloned downstream the RBS_B0034.</br> |
</br> | </br> | ||
We also cloned the T5LacOperator inside the pSB1C3 plasmid, making it ready for the submission. | We also cloned the T5LacOperator inside the pSB1C3 plasmid, making it ready for the submission. | ||
Revision as of 16:49, 23 September 2012
The Jolly JoCare
a safe probiotic platform for protein expression
Week 2
More
Suicide System
Finally we got by synthesis the antimicrobial peptide LL37 cathelicidin (BBa_K875009) in pEX-A plasmid and we amplified it through the transformation of DH5-alpha bacteria. The positive colonies were then digested EcoRI/SpeI, the fragment was eluted from the gel and then cut it again with EcoRI, because the pEX-A plasmid has another EcoRI restriction site downstream our fragment. Then we did a column purification of the fragment and we successfully cloned the LL 37 into the pSB1C3 linearized backbone to make it ready for a potential submission. We LL 37 -previously cut EcoRI/SpeI- was cloned upstream the TT_B0015 and the DH5alpha cells were transformed with the plasmid (LL-37-B0015). The positive colonies were inoculated and then cloned downstream the RBS_B0034. We also cloned the T5LacOperator inside the pSB1C3 plasmid, making it ready for the submission.Antibody
Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.Chassis
Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.
