Difference between revisions of "Team:Pasteur Paris/Description"
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<h4>BBa_K2053002 (for Silver medal) : Fusion Si4-cellulose-binding domain-B domain protein A</h4> | <h4>BBa_K2053002 (for Silver medal) : Fusion Si4-cellulose-binding domain-B domain protein A</h4> | ||
| − | In order to functionalize a cellulose-based patch into a biodetection device we designed and created a fusion protein by assembling three parts: the phage displayed silica-binding peptide (Si4) [3], the Clostridium cellulovorans cellulose-binding domain of cellulose-binding protein A (CBPa) [4], and the B domain of Staphylococcus aureus protein A (BpA) [5]. This fusion protein is used to bind to cellulose-based patch, increase its rigidity by biosilification, and make it a detection device by fixing Fc fragment of specific antibodies. We used preexisting iGEM BioBrick parts that we assembled thanks to commonly used flexible linkers [6]. </br></br> | + | In order to functionalize a cellulose-based patch into a biodetection device we designed and created a fusion protein by assembling three parts: the phage displayed silica-binding peptide (Si4) [3], the <i>Clostridium cellulovorans</i> cellulose-binding domain of cellulose-binding protein A (CBPa) [4], and the B domain of <i>Staphylococcus aureus</i> protein A (BpA) [5]. This fusion protein is used to bind to cellulose-based patch, increase its rigidity by biosilification, and make it a detection device by fixing Fc fragment of specific antibodies. We used preexisting iGEM BioBrick parts that we assembled thanks to commonly used flexible linkers [6]. </br></br> |
To facilitate protein purification, we added a HisTag. To remove the HisTag, we separated the HisTag from the sequence by the TEV cleavage site sequence. </br></br> | To facilitate protein purification, we added a HisTag. To remove the HisTag, we separated the HisTag from the sequence by the TEV cleavage site sequence. </br></br> | ||
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<h5>BBa_K2053003(for Gold medal) : Fusion cellulose-binding domain of CBP-B domain of protein A</h5> | <h5>BBa_K2053003(for Gold medal) : Fusion cellulose-binding domain of CBP-B domain of protein A</h5> | ||
| − | We improved previously existing BBa_K863110 and BBa_K103003 BioBricks parts by combining them, creating one part. It is a fusion protein between the Clostridium cellulovorans cellulose-binding domain of cellulose-binding protein A (CBPa) with the B domain of Staphylococcus aureus protein A (BpA). This fusion protein can be used to bind antibodies to cellulose.</br></br> | + | We improved previously existing BBa_K863110 and BBa_K103003 BioBricks parts by combining them, creating one part. It is a fusion protein between the <i>Clostridium cellulovorans</i> cellulose-binding domain of cellulose-binding protein A (CBPa) with the B domain of <i>Staphylococcus aureus</i> protein A (BpA). This fusion protein can be used to bind antibodies to cellulose.</br></br> |
To facilitate protein purification, we added a HisTag. To remove the HisTag, we separated the HisTag from the sequence by the TEV cleavage site sequence.</br> | To facilitate protein purification, we added a HisTag. To remove the HisTag, we separated the HisTag from the sequence by the TEV cleavage site sequence.</br> | ||
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<h2>References: </h2> | <h2>References: </h2> | ||
| − | [1] Characterization of an Endoplasmic Reticulum-associated Silaffin Kinase from the Diatom Thalassiosira pseudonana, Sheppard V et al, 2010, The Journal of Biological Chemistry</br> | + | [1] Characterization of an Endoplasmic Reticulum-associated Silaffin Kinase from the Diatom <i>Thalassiosira pseudonana</i>, Sheppard V et al, 2010, The Journal of Biological Chemistry</br> |
[2] The Role of Proteins in Biosilification, Otzen D, 2012, Scientifica</br> | [2] The Role of Proteins in Biosilification, Otzen D, 2012, Scientifica</br> | ||
[3] Silica-Precipitating Peptides Isolated from a Combinatorial Phage Display Peptide Library Naik et al, 2002, Journal of Nanoscience and Nanotechnology</br> | [3] Silica-Precipitating Peptides Isolated from a Combinatorial Phage Display Peptide Library Naik et al, 2002, Journal of Nanoscience and Nanotechnology</br> | ||
| − | [4] Characterization of the cellulose-binding domain of the Clostridium cellulovorans cellulose-binding protein A, Goldstein MA, 1993, J Bacteriol</br> | + | [4] Characterization of the cellulose-binding domain of the <i>Clostridium cellulovorans</i> cellulose-binding protein A, Goldstein MA, 1993, J Bacteriol</br> |
| − | [5] Crystallographic refinement and atomic models of a human Fc fragment and its complex with fragment B of protein A from Staphylococcus aureus at 2.9- and 2.8-A resolution, Deisenhofer J, 1981, Biochemistry</br> | + | [5] Crystallographic refinement and atomic models of a human Fc fragment and its complex with fragment B of protein A from <i>Staphylococcus aureus</i> at 2.9- and 2.8-A resolution, Deisenhofer J, 1981, Biochemistry</br> |
[6] Fusion Protein Linkers: Property, Design and Functionality, Xiaoying C, 2013, Advanced Drug Delivery Reviews</br> | [6] Fusion Protein Linkers: Property, Design and Functionality, Xiaoying C, 2013, Advanced Drug Delivery Reviews</br> | ||
