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Team:LMU-Munich/Results
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=== The SdpI BioBrick can rescue ∆''sdpI B. subtilis'' mutant strains === | === The SdpI BioBrick can rescue ∆''sdpI B. subtilis'' mutant strains === | ||
| - | + | Induceable SdpI production leads to higher resistance against the antimicrobial peptide SdpC. | |
[https://2014.igem.org/Team:LMU-Munich/Project/Bakillus#suicide (Further Information)] | [https://2014.igem.org/Team:LMU-Munich/Project/Bakillus#suicide (Further Information)] | ||
Revision as of 03:36, 18 October 2014
Results
Here we provide you with a quick overview of our key results. Please follow the links to receive more background information.
Sensing of Staphylococcus aureus
After coming across some decent issues with induction of the AgrC-IIA two-component system with synthetic AIP-II, and excluding a variety of causes, we identified extracellular proteases as possible cause for the problem. We successfully transformed the protease deficient strain B. subtilis WB700 with the reporter construct pBS4S-PxylA-agrCA. The next step will be the transformation with the reporter constructs pBS3C-P2/P3-luxABCDE.
Sensing of Streptococcus pneumoniae
The two-component system ComDE from Streptococcus pneumoniae, introduced into B. subtilis, can sense the QS-peptide CSP and activate the corresponding promoters PcomC and PcomAB.
Adhesion to Streptococcus pneumoniae
Three peptides were shown to bind to S. pneumoniae when fused to a maltose binding protein. Anchors to the B. subtilis cell wall are still in work.
Binding of the peptides C4P, CSP and A5P to S. pneumoniae cells was evaluated with the results strongly indicating the adhesion. Our BioBricks [http://parts.igem.org/Part:BBa_K1351001 BBa_K1351001], [http://parts.igem.org/Part:BBa_K1351002 BBa_K1351002] and [http://parts.igem.org/Part:BBa_K1351003 BBa_K1351003] thus can serve as a helpful tool for biomedical applications targeting S. pneumoniae.
Subtilin has inhibitory impact on S. pneumoniae
Spot-on-lawn assays proved the inhibitory effect of the antimicrobial agent on S. pneumoniae.
Subtilin has inhibitory impact on S. aureus
Spot-on-lawn assays proved the inhibitory effect of the antimicrobial agent on S. aureus, including MRSA types. Experiments were performed by our collaboration partner iGEM Team Groningen.
SpaIFEG immunity BioBrick ([http://parts.igem.org/Part:BBa_K1351014 BBa K1351014]) mediates self-protection from subtilin.
These amazing results were evaluated by subtilin-containing supernatant that was filled in holes of agar plates and thus inhibited the growth of the B. subtilis wild type but inhibited the growth of the B. subtilis W168 amyE::Pxyl-spaIFEG significantly less.
Ecf41 provides a delayed expression
A delayed expression can be achieved by wiring the ecf σ factor σecf41 and its target promoter PydfG between the input promoter and the output.
The SdpI BioBrick can rescue ∆sdpI B. subtilis mutant strains
Induceable SdpI production leads to higher resistance against the antimicrobial peptide SdpC.
The sdpI-antisense RNA shows no impact regarding accelerated degradation
In B. subtilis W168 cells the expression of the sdpI-antisense RNA did not show any impact on cell vitality.
BioBrickBox
To extend the Bacillus BioBrick Box started by the 2012 team LMU-Munich, we created a library of ribosome binding sites and protein linkers for B. subtilis.
Modeling
We determined the influence of relevant parameters to the dynamic of the induced inhibition of the immunity protein production for the suicide switch. We now can tell which parameters benefit the suicide and which are in reciprocal relation.
Furthermore, we were able to simulate the chemotactical driven movement of BaKillus according to a given concentration gradient of respective substances.
Application
We developed a nasal spray for the application of BaKillus against sinusitis.(Further Information)
Furthermore we checked in a self-experiment that B.subtilis is viable in the nasal mucosa and can survive long enough to conduct its job of killing pathogens.
Team LMU-Munich
igem@bio.lmu.de
AG Thorsten MascherGroßhaderner Str. 2
82152 Planegg (Martinsried)
Hi there!
Welcome to our Wiki! I'm BaKillus, the pathogen-hunting microbe, and I'll guide you on this tour through our project. If you want to learn more about a specific step, you can simply close the tour and come back to it anytime you like. So let's start!
What's the problem?
First of all, what am I doing here? The problem is, pathogenic bacteria all around the world are becoming more and more resistant against antimicrobial drugs. One major reason for the trend is the inappropriate use of drugs. With my BaKillus super powers, I want to reduce this misuse and thus do my part to save global health.
Sensing of pathogens
To combat the pathogenic bacteria, I simply eavesdrop on their communication. Bacteria talk with each other via quorum sensing systems, which I use to detect them and trigger my responses.
Adhesion
The more specific and effective I can use my powers, the lower the danger is of provoking new resistance development. So I catch pathogens whenever I get hold of them and stick to them until my work is done.
Killing
Talking about my work - killing pathogens is finally what I am made for. In response to quorum sensing molecules of the pathogens, I export a range of antimicrobial substances leading to dissipation of biofilms and the killing of the targeted bacteria.
Suicide switch
When the job is done and all the bad guys are finished, you don't need a super hero anymore. So after fulfilling my work I say goodbye to the world by activating my suicide switch.
Application
Of course I'm not only a fictional hero, but a very real one. In two different prototypes, I could be used for diagnosis or treatment of pathogen-caused diseases. However, there is still a whole lot of regulational and economical questions that have to be answered before.
See you!
So now you know my short story - and it is time for me to return to my fight for a safer world. Feel free to take a closer look on my super powers, the process of my development or the plans for a medical application.
