Difference between revisions of "Team:Nankai/Safety"

 
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                    <p>Your place:&nbsp;<a href="https://2015.igem.org/Team:Nankai">Home</a>&nbsp;&gt;&nbsp;<a href="https://2015.igem.org/Team:Nankai/Description">Project</a>>&nbsp;&gt;&nbsp;<a href="https://2015.igem.org/Team:Nankai/Safety">Safety</a></p>
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                  <p>Your place:&nbsp;<a href="https://2015.igem.org/Team:Nankai">Home</a>&nbsp;&gt;&nbsp;<a href="https://2015.igem.org/Team:Nankai/Safety">Safety</a></p>
 
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<h2 class="page-title">Project Safety</h2>
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<h4>What is γ-PGA?</h4>
 
<p>Poly-γ-glutamic acid (γ-PGA) is an important, naturally occurring polyamide consisting of D/L-glutamate monomers. Unlike typical peptide linkages, the amide linkages in γ-PGA are formed between the α-amino group and the γ-carboxyl group. γ-PGA exhibits many favorable features such as biodegradable, water soluble, edible and non-toxic to humans and the environment. Therefore, it has been widely used in fields of foods, medicines, cosmetics and agriculture and many unique applications, such as a sustained release material and drug carrier, curable biological adhesive, biodegradable fibres, and highly water absorbable hydrogels.</br></p>
 
<h4>How can we produce it?</h4>
 
<p>Strains capable for producing γ-PGA are divided into two categories based on their requirement for glutamate acid: glutamate-dependent strains and glutamate-independent strains. Glutamate-independent strains are preferable for industrial production because of their low cost and simplified fermentation process. However, compared with glutamate-dependent strains, their lower γ-PGA productivity limits their industrial application. Therefore, the construction of a glutamate-independent strain with high γ-PGA yield is important for industrial applications.</br></p>
 
<h4>Who can produce it?</h4>
 
<p>Bacillusamyloliquefaciens LL3, isolated from fermented food, is a glutamate-independent strain, which can produce 3-4 g/L γ-PGA with sucrose as its carbon source and ammonium sulfate as its nitrogen source. The B. amyloliquefaciens LL3 strain was deposited in the China Center for Type Culture Collection (CCTCC) with accession number CCTCC M 208109 and its whole genome has been sequenced in 2011. In this study, we aimed to improve the γ-PGA production based on the B. amyloliquefaciens NK-1 strain (a derivative of LL3 strain with its endogenous plasmid and upp gene deleted).</br></p>
 
<h4>What did we do?</h4>
 
<p>In order to improve γ-PGA production, we employed two strategies to fine-tune the synthetic pathways and balance the metabolism in the glutamate-independent B. amyloliquefaciens NK-1 strain. Firstly, we constructed a metabolic toggle switch in the NK-1 strain to inhibit the expression of ODHC (2-oxoglutarate dehydrogenase complex) by adding IPTG in the stationary stage and distribute the metabolic flux more frequently to be used for γ-PGA precursor-glutamate synthesis. As scientists had found that the activity of ODHC was rather low when glutamate was highly produced in a Corynebacterium glutamicum strain. Second, to balance the increase of endogenous glutamate production, we optimized the expression level of pgsBCA genes (responsible for γ-PGA synthesis) by replacing its native promoter to seven different strength of promoters. Through these two strategies, we aimed to obtain a γ-PGA production improved mutant strain.<a href="https://2015.igem.org/Team:Nankai/Experiments">Click for more detail.</a></p>
 
 
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<h4>GENERAL SAFETY</h4>
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<p>DEALING with bacteria is never a work without risk, and all our members are fully aware of this. We have done a lot of work on safety. These work mainly contain:
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<strong>1)</strong>adequate training for every member who is going to participate in laboratory work;
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<strong>2)</strong>consulting with directors to make sure our job is safe;                                                                             
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<strong>3)</strong>communication with local safety affair office;
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<strong>4)</strong>All the organisms we use in our lab are safety level 1 organisms. We only operate on those bacteria in clean benches and all the waste contains microorganisms are sterilized before further dispose. During experiments we  rubber gloves, disposable masks and goggles if it is necessary.</p>
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<h4>SAFE TEAM MEMBERS ^-^</h4>
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<p>WE'RE highly aware that we ourselves are the most important forces to avoid all the possible risks, as well as the most possible devils to cause all the disasters. So we pay much much attention on relative training to ensure that our members know clearly what should do and what should not in laboratory.</p>
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<h4>SAFE DESIGN</h4>
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<p>PROJECT design is one of the most well-thought factors in our work. We have some members in our team whose work is specifically investigate the design made by those members who undertake laboratory works to make sure there is no obviously risky items. <strong>All</strong> those investigated design will be later sent to our instructor to be double checked. </p>
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<h4>SAFE LAB WORK</h4>
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<p>LAB WORK is the most most important part of safety, and we are strictly careful on this. Usually when we carry out our lab work, we will choose some members to inspect those carriers to make sure their work is safe.And all our waste are discarded safely, strictly followed stipulations of our school. </p>
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<h4>SAFE MATERIALS</h4>
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<p>THE BACTERIA we choose is a <em>Bacillus amyloliquefaciens</em> strain we isolated from fermented food. It's hard to directly operate on this Bacillus amyloliquefaciens strain genetically. So first we constructed expression vectors in <em>E. coli</em> DH5alpha, demethylated the plasmids in <em>E. coli</em> GM110, and then used methylase extracted from Bacillus amyloliquefaciens to methylated the plasmids. At last we transformed those functional plasmids into Bacillus. ALL THE ORGANISM we use in our lab are safety level 1 organisms. We only operate on those bacteria in clean benches and all the waste contains microorganisms are sterilized before further dispose. During experiments we  rubber gloves, disposable masks and goggles if it is necessary.</p>
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<h5 class="widget-title"><a href="https://2015.igem.org/Team:Nankai/Description">Description</a></h5>
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                                                <p>SAFETY is highly taken care of HERE!</p>
<h5 class="widget-title"><a href="https://2015.igem.org/Team:Nankai/Experiments">Experiments & Protocols</a></h5>
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        <img src="https://static.igem.org/mediawiki/2015/0/03/Nankai_Safetypic2.jpg">
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                                                <p>We ONLY work in a tidy lab!</p>
<h5 class="widget-title"><a href="https://2015.igem.org/Team:Nankai/Results">Results</a></h5>
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<h5 class="widget-title"><a href="https://2015.igem.org/Team:Nankai/Design">Design</a></h5>
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                                                <p>We are STRICT with our products!.</p>
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                                                <p>When working,we ALWAYS have someone to INSPECT!</p>
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Latest revision as of 14:55, 18 September 2015

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Your place: Home > Safety

Safety

GENERAL SAFETY

DEALING with bacteria is never a work without risk, and all our members are fully aware of this. We have done a lot of work on safety. These work mainly contain: 1)adequate training for every member who is going to participate in laboratory work; 2)consulting with directors to make sure our job is safe; 3)communication with local safety affair office; 4)All the organisms we use in our lab are safety level 1 organisms. We only operate on those bacteria in clean benches and all the waste contains microorganisms are sterilized before further dispose. During experiments we rubber gloves, disposable masks and goggles if it is necessary.

SAFE TEAM MEMBERS ^-^

WE'RE highly aware that we ourselves are the most important forces to avoid all the possible risks, as well as the most possible devils to cause all the disasters. So we pay much much attention on relative training to ensure that our members know clearly what should do and what should not in laboratory.

SAFE DESIGN

PROJECT design is one of the most well-thought factors in our work. We have some members in our team whose work is specifically investigate the design made by those members who undertake laboratory works to make sure there is no obviously risky items. All those investigated design will be later sent to our instructor to be double checked.

SAFE LAB WORK

LAB WORK is the most most important part of safety, and we are strictly careful on this. Usually when we carry out our lab work, we will choose some members to inspect those carriers to make sure their work is safe.And all our waste are discarded safely, strictly followed stipulations of our school.

SAFE MATERIALS

THE BACTERIA we choose is a Bacillus amyloliquefaciens strain we isolated from fermented food. It's hard to directly operate on this Bacillus amyloliquefaciens strain genetically. So first we constructed expression vectors in E. coli DH5alpha, demethylated the plasmids in E. coli GM110, and then used methylase extracted from Bacillus amyloliquefaciens to methylated the plasmids. At last we transformed those functional plasmids into Bacillus. ALL THE ORGANISM we use in our lab are safety level 1 organisms. We only operate on those bacteria in clean benches and all the waste contains microorganisms are sterilized before further dispose. During experiments we rubber gloves, disposable masks and goggles if it is necessary.

 

Medigo Blue, free responsive template