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Team:StanfordBrownSpelman/Cellulose Cross Linker
From 2014.igem.org
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| - | <div class="sub4"><a href=" | + | <div class="sub4"><a href="http://docs.google.com/document/d/1-BS2AXdxk_gbYPC2qc5T6e1L7qtnXg4DxsOF-aoxIQg/edit?usp=sharing" target="_blank"><img src="https://static.igem.org/mediawiki/2014/2/25/SBS_iGEM_2014_download.png"></a><a href="http://docs.google.com/document/d/1-BS2AXdxk_gbYPC2qc5T6e1L7qtnXg4DxsOF-aoxIQg/edit?usp=sharing">Click here to go to our project journal, which details our design and engineering process and included descriptions of the protocols we developed and used.</a></div> |
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Revision as of 05:44, 17 October 2014
Cellulose Cross-Linker
The goal of this subproject is to create a cellulose cross-linking protein to increase material strength and allow for the modular attachment of biological sensors. This fusion protein contains two distinct cellulose-binding domains[1] on either side of a streptavidin domain. The cellulose-binding domains cross link the cellulose fibers while the streptavidin serves as a binding domain for biological sensors. The interaction between SA (streptavidin) and biotin is one of the strongest non-covalent interactions in nature [2]. Therefore a cell expressing an outer membrane protein that has been biotinylated will bind tightly to this domain. This will allow our UAV to make use of a number of biological sensors.
The goal of this subproject is to create a cellulose cross-linking protein to increase material strength and allow for the modular attachment of biological sensors. This fusion protein contains two distinct cellulose-binding domains[1] on either side of a streptavidin domain. The cellulose-binding domains cross link the cellulose fibers while the streptavidin serves as a binding domain for biological sensors. The interaction between SA (streptavidin) and biotin is one of the strongest non-covalent interactions in nature [2]. Therefore a cell expressing an outer membrane protein that has been biotinylated will bind tightly to this domain. This will allow our UAV to make use of a number of biological sensors.
Approach & Methods
Figure 1. An illustration of cellulose binding domains cross-linking cellulose fibers with a streptavidin domain in the middle. The biosensing cell is expressing a biotinylated AviTag which will bind to the streptavidin .
Results
Our initial approach was to include two identical cellulose-binding domains on either side of the streptavidin domain. Due to the repetitive nature of the sequence and potential homologous recombination, we had many issues with molecular cloning. We changed our approach to using two different cellulose-binding domains with different sequences. This allowed us to successfully conduct the molecular cloning.
Figure 2. Sequencing data for the cross-linking protein. The solid green bar indicates a perfect match between our sequence and the expected sequence.The first 1000 base pairs are sequenced in this forward sequence.
