Difference between revisions of "Team:Pasteur Paris/Safety Security"
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The following <a href="http://parts.igem.org/Part:BBa_K2053002">Biobrick BBa_K2053002</a> has been characterized thoroughly by experiments. </br> | The following <a href="http://parts.igem.org/Part:BBa_K2053002">Biobrick BBa_K2053002</a> has been characterized thoroughly by experiments. </br> | ||
| − | This fusion protein is at the heart of our patch. It combines Silification, cellulose binding and anti-body fixation. Please follow our project design <a href="https://2016.igem.org/Team:Pasteur_Paris/Science">Science page</a> for details. </br></br></br> | + | This fusion protein is at the heart of our patch. It combines Silification, cellulose binding and anti-body fixation. Please follow our project design <a href="https://2016.igem.org/Team:Pasteur_Paris/Science">Science page</a> for details about it's properties, and it's safety compliance are described below. </br></br></br> |
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<h2><B>Protein domain functions</B></h2> | <h2><B>Protein domain functions</B></h2> | ||
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| − | • In <i>Staphylococcus aureus</i>, the protein A (BpA) binds to constant fragments of IgG antibodies in order to inhibit opsonophagocytosis, and might play a role as an adhesin during the initiation of intravascular infection by mediating attachment of <i>S. aureus</i> to proteins at the site of damage to the endothelium. This protein is neither a toxin nor an enzyme that synthesizes a toxin. This protein is a staphylococcal virulence factor involved in immune evasion and adherence of bacteria onto endothelium. In order to make an immunodetection-based vector-borne pathogens sensor, we will use this part as a binder for specific antibodies. We do not require the whole protein, thus we will only use the antibody-binding domain of this protein: the B domain. Based on the registry part sequence, we will perform gblock oligonucleotide synthesis (iGEM pre-existing part: <a href="http://parts.igem.org/Part:BBa_K103003">BBa_K103003</a>).</br></br> | + | • In <i>Staphylococcus aureus</i>, the protein A (BpA) binds to constant fragments of IgG antibodies in order to inhibit opsonophagocytosis, and might play a role as an adhesin during the initiation of intravascular infection by mediating attachment of <i>S. aureus</i> to proteins at the site of damage to the endothelium. <B>This protein is neither a toxin nor an enzyme that synthesizes a toxin</B>. This protein is a staphylococcal virulence factor involved in immune evasion and adherence of bacteria onto the endothelium. In order to make an immunodetection-based vector-borne pathogens sensor, we will use this part as a binder for specific antibodies. We do not require the whole protein, thus we will <B>only use the antibody-binding domain of this protein</B>: the B domain. Based on the registry part sequence, we will perform gblock oligonucleotide synthesis (iGEM pre-existing part: <a href="http://parts.igem.org/Part:BBa_K103003">BBa_K103003</a>).</br></br> |
| − | • The cellulose-binding protein is secreted by <i>Clostridium cellulovorans</i> to bind cellulose fibers and coordinate cellulase enzymes. The catabolic process of cellulose by this bacterium results in increase of glucose, an essential nutrient for bacterium growth. This protein is neither a toxin nor an enzyme that synthesizes a toxin. Moreover, the coordination of cellulose enzymes by this protein is not a problem, because these enzymes are absent in humans. In order to make a cellulose-based vector-borne pathogens sensor, we will use this part as a binder of a cellulose matrix. We do not require the whole protein, that is why we will only use the cellulose-binding domain of this protein. Based on the registry part sequence, we will perform gblock oligonucleotide synthesis (iGEM pre-existing part: <a href="http://parts.igem.org/Part:BBa_K863110">BBa_K863110</a>).</br></br> | + | • The cellulose-binding protein is secreted by <i>Clostridium cellulovorans</i> to bind cellulose fibers and coordinate cellulase enzymes. The catabolic process of cellulose by this bacterium results in increase of glucose, an essential nutrient for bacterium growth. <B>This protein is neither a toxin nor an enzyme that synthesizes a toxin</B>. Moreover, the coordination of cellulose enzymes by this protein is not a problem, because these enzymes are absent in humans. In order to make a cellulose-based vector-borne pathogens sensor, we will use this part as a binder of a cellulose matrix. We do not require the whole protein, that is why we will only use the cellulose-binding domain of this protein. Based on the registry part sequence, we will perform gblock oligonucleotide synthesis (iGEM pre-existing part: <a href="http://parts.igem.org/Part:BBa_K863110">BBa_K863110</a>).</br></br> |
| − | • The silica-binding domain from phage screen is able to condense silicic acid to form biosilica. This protein is neither a toxin nor an enzyme that | + | • The silica-binding domain from phage screen is able to condense silicic acid to form biosilica. <B>This protein is neither a toxin nor an enzyme that synthesizes a toxin</B>. In order to make a silica-based vector-borne pathogens sensor, we will use this part as a binder of silicic acid to enhance biosilica formation and increase the rigidity of the cellulose-base matrix. Based on the registry part sequence, we will perform gblock oligonucleotide synthesis (iGEM pre-existing part: <a href="http://parts.igem.org/Part:BBa_K1028000">BBa_K1028000</a>).</br></br> |
| − | • Since the silica-binding peptide (Si4), the cellulose-binding domain of cellulose-binding protein (CBPa), and the B domain of staphylococcal protein A are individually non toxic, we assume that the combination of them (newly iGEM 2016 deposited part: <a href="http://parts.igem.org/Part:BBa_K2053000">BBa_K2053002</a>) will also be non toxic, although this remains to be tested in the lab.</br></br> | + | • Since the silica-binding peptide (Si4), the cellulose-binding domain of cellulose-binding protein (CBPa), and the B domain of staphylococcal protein A are individually non toxic, we assume that the <B>combination</B> of them (newly iGEM 2016 deposited part: <a href="http://parts.igem.org/Part:BBa_K2053000">BBa_K2053002</a>) will <B>also be non toxic</B>, although this remains to be tested in the lab.</br></br> |
| − | • The silafin kinase TpsTK1 (newly iGEM 2016 deposited part: <a href="http://parts.igem.org/Part:BBa_K2053000">BBa_K2053000</a> is involved in the biomineralization of silafins. They are quite abundant in the diatoms, and help in enhancing the phosphorylation of the silificating peptides.There is no bibliographic evidence of toxicity (Pubmed, Google search). </br></br> | + | • The silafin kinase TpsTK1 (newly iGEM 2016 deposited part: <a href="http://parts.igem.org/Part:BBa_K2053000">BBa_K2053000</a> is involved in the biomineralization of silafins. They are quite abundant in the diatoms, and help in enhancing the phosphorylation of the silificating peptides.<B>There is no bibliographic evidence of toxicity</B> (Pubmed, Google search). </br></br> |
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| − | These components used in our experiments are classified as <B>biosafety level 1</B> organisms; all are harmless and were used under well-established protocols and with proper guidance and safety equipment. The safety levels of our laboratory are BHSL1 and 2. These biosafety levels are suitable for work involving potentially biohazardous agents to personnel and the environment. </br> | + | These components used in our experiments are classified as <B>biosafety level 1 (BHSL 1)</B> organisms; all are harmless and were used under well-established protocols and with proper guidance and safety equipment. The safety levels of our laboratory are BHSL1 and 2. These biosafety levels are suitable for work involving potentially biohazardous agents to personnel and the environment.</br> |
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<img src="https://static.igem.org/mediawiki/2016/7/77/Safety1_pasteur.png" width="42%" alt="image"/></img> | <img src="https://static.igem.org/mediawiki/2016/7/77/Safety1_pasteur.png" width="42%" alt="image"/></img> | ||
| − | <B>All lab work and experiments were done according to laboratory safety policy of the Institut Pasteur</B>. Students | + | <B>All lab work and experiments were done according to laboratory safety policy of the Institut Pasteur</B>. Students were taught a course on common hazard and exposure risks in the lab, including chemical and biological hazards that are found at the Institut Pasteur research laboratories. The course taught us how to prevent exposure to these hazards and emergency response procedures in case of exposure. The course also covered our lab waste handling procedures as well as useful information on methods to ensure the research laboratory is free from common physical hazards. The training was performed by Dr Deshmukh Gopaul following guidelines from the Health and safety department of Institut Pasteur. </br></br></br> |
| − | <B>N.B:</B> Since not all team members have had wet lab experience before, all the lab work was done under the supervision and guidance of at least one of our authorized senior team members and/or advisors. Secondly, the | + | <B>N.B:</B> Since not all team members have had wet lab experience before, all the lab work was done under the supervision and guidance of at least one of our authorized senior team members and/or advisors. Secondly, the immuno-detection tests that required experiments with infected mosquitoes were conducted by one of our coaches, who has clearance for this level, in a biosafety level 3 laboratory. Mosquitoes, rated pathogen-free, involved in the other experiments came from an insectarium in Institut Pasteur, none were taken from the environment. All wastes were treated according to the disposal routes established for decontamination, and autoclaving</br></br> |
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All lab work and experiments were done according to laboratory safety policy of the Institut Pasteur. Students were taught a course on common hazard and exposure risks in the lab, including chemical and biological hazards that are found at the Institut Pasteur research laboratories. The course taught us how to prevent exposure to these hazards and emergency response procedures in case of exposure. The course also covered our lab waste handling procedures as well as useful information on methods to ensure the research laboratory is free from common physical hazards. The training was performed by Dr Deshmukh Gopaul following guidelines from the Health and safety department of Institut Pasteur.
N.B: Since not all team members have had wet lab experience before, all the lab work was done under the supervision and guidance of at least one of our authorized senior team members and/or advisors. Secondly, the immuno-detection tests that required experiments with infected mosquitoes were conducted by one of our coaches, who has clearance for this level, in a biosafety level 3 laboratory. Mosquitoes, rated pathogen-free, involved in the other experiments came from an insectarium in Institut Pasteur, none were taken from the environment. All wastes were treated according to the disposal routes established for decontamination, and autoclaving
