Team:Trieste/data

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Data

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An overview on our system



This is a diagram summarising our system. It consists of two parts:the production of Ab against NoroVirus and the gene guard.
Concerning the gene guard, CymR inhibits the T5 CUmate Operator avoiding the production of the T4 Holin (permeabilizes the cytoplasmatic membrane) and the LL 37 cathelicidin (human antimicrobial peptide able to distrupt the outer membrane).
When the cumate is given, it binds and inactivates the CymR, derepressing the T5 Cumate Operator Promoter and allowing the expression of T4 Holin, that permeabilizes the cytoplasmatic membrane, allowing the secretion of LL 37.

Our favourite parts:




BBa_K875001, T5 cumate operator:


We have dimontrated that it is a very strong and strictly regulated promoter. Thanks to its rapid reactivity it's one of the first candidate for the expression and production of toxin in case of necessity.

BBa_K875003, CymR:


We have succesfully expressed the CymR. We have also showed his stringency as repressor of the T5 Cumate Operator that makes the all system a very useful mechanism for an high specificity expression.

BBa_K875004, LPP-OmpA-scFv:


We have verified the production of this protein at high yield. OmpA is a very powerful method for displaying proteins on the external membrane, in this case the antibody scFv 54.6.

The part we have improved:

BBa_K875020, Beta-Glucosidase (Osaka 2010):


WThis composite part is developed by previous work by UNITS iGEM team 2011 and Edinburgh team 2011. We improved and characterized this part demonstrating its efficiency.


Other parts that we have developed:


BBa_K875002, T5 Lac Operator:


We have shown that this promoter is a very useful biobricks in the primary testing of protein expression thanks to his handiness.

BBa_K875005, LPP-OmpA-SIP:


We have built this biobrick that is small and very stable at the same time. Thank to this high stability it is a valid alternative to the LPP-OmpA-scFv.

BBa_K875006, PelB-scFv:


We have create this composit as an optimum system to send the protein inside the bacterial periplasm and also outside the outer mambrane according to the protein size.

BBa_K875007, PelB-SIP:


We have built this biobrick because it is known from literature that the secreted SIP acts again enteric infection.

BBa_K875008, Tse2 toxin:


We demonstrate that this composit is a important way to affect the bacterial life intracellularly.

BBa_K875009, LL37-T4 holin:


We projected this composit using LL-37 which is a very powerful human antimicrobial peptide that kills bacteria destroying the outer membrane. This toxin acts in combination with the T4 holin which breaks the inner membrane allowing the contact between LL-37 and the outer membrane.
Team iGEM 2012

Contact us

For other information, write to:

igem2012@gmail.com
Università degli studi di Trieste ICGEB Illy Fondazione Cassa di Risparmio
iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012
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