Difference between revisions of "Team:Pasteur Paris/Description"
| Line 114: | Line 114: | ||
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> | ||
| − | <h4>Fusion Si4-cellulose-binding domain-B of domain protein A | + | <h4>BBa_K2053002 (for Silver medal) : Fusion Si4-cellulose-binding domain-B of 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 <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 the Fc fragment of specific antibodies. We used the 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 the Fc fragment of specific antibodies. We used the preexisting iGEM BioBrick parts that we assembled thanks to commonly used flexible linkers [6]. </br></br> | ||
| Line 120: | Line 120: | ||
To facilitate protein purification, we added a His-Tag composed of 6 histidines at the N-terminus, or C-terminus. To remove the His-Tag, we inserted a TEV (Tobacco Etch virus) cleavage site downstream from the His-Tag. </br></br> | To facilitate protein purification, we added a His-Tag composed of 6 histidines at the N-terminus, or C-terminus. To remove the His-Tag, we inserted a TEV (Tobacco Etch virus) cleavage site downstream from the His-Tag. </br></br> | ||
| − | <h5> | + | <h5> Fusion cellulose-binding domain of CBP-B domain of protein A (for Gold medal)</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 <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> | 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> | ||
