Team:Leeds/Parts

From 2013.igem.org

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Please check back here frequently for a full list of the parts we've developed, submitted to the [http://partsregistry.org Registry] and have had approved. We will also endeavour to include our characeterisation protocols, although we are currently collaborating with [[Team:Purdue|Purdue]] to decide exactly what these should be.
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==Parts Summary==
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* [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1028000 Silica Binding Peptide (Si4):] A short amino acid sequence that binds silica with high affinity.
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'''Note to Team:''' ''proper part documentation is to describe and define a part such that it can be used without a need to refer to the primary literature. The next iGEM team should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for  users who wish to know more''
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* [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1028002 Bio-system 1:] CpxR promoter controlling GFP expression. Will produce the green fluorescent protein following exposure to hydrophobic membrane stress. Extensive experimental data has been acquired about the response of the cpxR promoter to various forms of physical and chemical stresses, this can be found in the Device 1 section of the Lab Results page.
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* [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1028003 Bio-system 2:] INP.Si4 is produced constitutively with GFP production under control of CpxR promoter. The system is designed to bind with high affinity to silica macro-structures and produce a fluorescent response when it does. Flow cytometry, fluorescence confocal, and scanning electron microscopy data has been gathered to characterize this part. Problems with expression meant that the data is not as conclusive as predicted but indicates that Si4 is expressed in some cells.
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==Currently Working On==
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* Bio-system 1: pCpxR promoter activates GFP expression. Will produce the green fluorescent protein on membrane stress.
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* Bio-system 2: Constitutive production of the Ice nucleation protein with a silica binding domain at the c-terminus.
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* Bio-system 3: integration of 1 and 2, INP Si4is constitutively produced but the fluorescent protein is under control of pCpxR promoter. When bound to silica beads will fluoresce
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* Also planning on having the silica binding peptide submitted as its own part for others to use for their own purposes.
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==Submitted==
==Submitted==
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<groupparts>iGEM013 Leeds</groupparts>
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==Characterisation Protocols==
 
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* Bio-system 1: Bacteria exposed to hydrophobic beads to induce membrane stress and fluorescence response is measured using a flourimeter.
 
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* Bio-system 2: Bacteria exposed to silica beads and washed. Control with no silica binding domain. Those that have bound will be visualised on the beads and those that have not will be seen in the elution. 
 
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* Bio-system 3: Expose the bacteria to silica beads will fluoresce when bound and not fluoresce when unbound. Measure the fluorescence with flourimeter.
 
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* The Si4 domain will be characterised with bio-system 2 if binding occurs.
 
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* Other characterisations we plan on doing are plate imaging, restriction mapping, growth curves, sequencing, and atomic force microscopy.
 
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Latest revision as of 03:51, 5 October 2013

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 ==Parts Summary==
  • Silica Binding Peptide (Si4): A short amino acid sequence that binds silica with high affinity.
  • Bio-system 1: CpxR promoter controlling GFP expression. Will produce the green fluorescent protein following exposure to hydrophobic membrane stress. Extensive experimental data has been acquired about the response of the cpxR promoter to various forms of physical and chemical stresses, this can be found in the Device 1 section of the Lab Results page.
  • Bio-system 2: INP.Si4 is produced constitutively with GFP production under control of CpxR promoter. The system is designed to bind with high affinity to silica macro-structures and produce a fluorescent response when it does. Flow cytometry, fluorescence confocal, and scanning electron microscopy data has been gathered to characterize this part. Problems with expression meant that the data is not as conclusive as predicted but indicates that Si4 is expressed in some cells.


Submitted

A full listing of all parts Leeds iGEM 2013 have submitted to the Registry, and their status.
<groupparts>iGEM013 Leeds</groupparts>

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Geneious, our fine sponsors and suppliers of software Bioline, our fine sponsors and suppliers of equipment Qiagen, our fine sponsors and suppliers of PCR kits
Bangs Laboratories, our fine sponsors and suppliers of silica beads
Leeds Homepage
Retrieved from "http://2013.igem.org/Team:Leeds/Parts"