Difference between revisions of "Team:SDSZ China"

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<p>Bovine milk, something so common that people consume every single day, inevitably contains antibiotics from the drugs or fodders that prevent them from mammary glands inflammation. Though strict protocols for diary products are enacted worldwide to make sure the drug residues are under the dosage that could lead to health problems for human, the current testing methods are not of great efficiency or accessibility, because of the high cost and complex procedures. This disadvantage leaves potential risks of letting milk containing antibiotics that are over the limit into the market.</p>
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        <h1 style="position:relative; font-family: Raleway; align-self: center;top:45%;font-size: 3em;"> TEST what you can't TASTE.</h1>
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Our project aims at speeding up while lowering the cost of the method used to detect antibiotic residues in milk. We take β-lactams as the target, and implement a penicillin binding protein, which recognizes all β-lactams, in our testing device. First, we improve the ELISA (enzyme linked immunosorbent assay) method, making the raw materials cheaper by replacing the immune response that occurs on the ELISA plate with a simple competitive binding reaction, in which the penicillin (or other β-lactams) on the plate and in the testing sample compete to bind with GFP-PBP5 (green fluorescent protein - penicillin binding protein 5). We can then measure the concentration of penicillin in the sample by measuring the fluorescence intensity on the plate. To even further reduce the budget and strengthen the sensitivity of the test, we created a testing paper by binding our fusion protein CBD (cellulose binding protein)-PBP5 to filter paper. When the paper is dipped in a milk sample, the presence of β-lactams alters PBP5’s enzymatic activity, which can be measured through electric conductivity test. Its electric conductivity therefore indicates how much of this type of antibiotic exists in our milk.
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So far, we have constructed pET-GFP-PBP5 and pET-CBD-PBP5. The plasmids are to be integrated into bacterial genes and express the fusion proteins that will be used in our device.  
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                    <a href="https://2016.igem.org/Team:SDSZ_China/Background"><p class="title"> Background</p>
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Our methodology provides new solutions for simplifying and reducing the cost of a standardized antibiotic residue testing procedure, making bovine milk safer to drink in China, and maybe someday throughout the world.
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                      Antibiotic injection is a common practice in dairy farms as a way to prevent mastitis in cow’s lactation period. But human’s long-term consumption of milk with excessive amount of antibiotics, can pose severe health threats.</p><br/>
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<p>In rural areas of China, milk goes directly to consumers without being tested for antibiotic residues, part of this is because currently used detection methods are either unreliable or not cost-effective enough to be accessed easily. Therefore we work to develop a cheap penicillin detection method, so that it can be used in relatively underdeveloped areas, and people in China could take one step closer to having an equal chance to qualified milk.
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                    <a href="https://2016.igem.org/Team:SDSZ_China/Integrated_Practices"><p class="title">Human Practice</p>
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                        To gain a better understanding of how our project can help improve the situation of antibiotic residues
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                        in China, we interviewed Dr. Meng Lu from Chinese Academy of Agricultural Science for more information,
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                        and visited the plant of Sanyuan Milk Group, one of China’s largest dairy processing companies.
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                        We integrated our research and outreach to the optimization of our project, hoping to lessen the
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                        inequality between rural and urban residents’ access to qualified milk.
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                    <p class="title">Project</p>
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                        <span class="subtitle">1.0</span> We engineer our bacteria to generate Penicillin Binding Protein 5-Green fluorescence Protein (PBP5-GFP), and bind penicillin to the coated wells on the ELISA plate. The protein is added to the sample, where the penicillin in the sample compete for PBP5-GFP with those on the plate, and the fluorescence intensity in the sample is measured and reflects its penicillin concentration.
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                        <span class="subtitle">2.0</span> PBP5 causes the hydrolysis of tripeptide Lys-Ala-Ala(KAA) when active and not bound with penicillin, releasing one alanine and exposing one amino group.[3] We measure its hydrolysis by the amino group, which changes the color of KAISER, a substance that reacts with free amino acids to yield CO2, NH3, and an aldehyde, the NH3 further yielding a colored product (diketohydrindylidene-diketohydrindamine, a bi-indanedione derivative also called Ruhemann purple), yielding the color purple. With more penicillin present, more PBP5 becomes inactive, and the purple gets lighter.
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Latest revision as of 20:19, 19 October 2016

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Test What You Can't TASTE

TEST what you can't TASTE.

Background


Antibiotic injection is a common practice in dairy farms as a way to prevent mastitis in cow’s lactation period. But human’s long-term consumption of milk with excessive amount of antibiotics, can pose severe health threats.


In rural areas of China, milk goes directly to consumers without being tested for antibiotic residues, part of this is because currently used detection methods are either unreliable or not cost-effective enough to be accessed easily. Therefore we work to develop a cheap penicillin detection method, so that it can be used in relatively underdeveloped areas, and people in China could take one step closer to having an equal chance to qualified milk.

Human Practice


To gain a better understanding of how our project can help improve the situation of antibiotic residues in China, we interviewed Dr. Meng Lu from Chinese Academy of Agricultural Science for more information, and visited the plant of Sanyuan Milk Group, one of China’s largest dairy processing companies. We integrated our research and outreach to the optimization of our project, hoping to lessen the inequality between rural and urban residents’ access to qualified milk.

Project


1.0 We engineer our bacteria to generate Penicillin Binding Protein 5-Green fluorescence Protein (PBP5-GFP), and bind penicillin to the coated wells on the ELISA plate. The protein is added to the sample, where the penicillin in the sample compete for PBP5-GFP with those on the plate, and the fluorescence intensity in the sample is measured and reflects its penicillin concentration.


2.0 PBP5 causes the hydrolysis of tripeptide Lys-Ala-Ala(KAA) when active and not bound with penicillin, releasing one alanine and exposing one amino group.[3] We measure its hydrolysis by the amino group, which changes the color of KAISER, a substance that reacts with free amino acids to yield CO2, NH3, and an aldehyde, the NH3 further yielding a colored product (diketohydrindylidene-diketohydrindamine, a bi-indanedione derivative also called Ruhemann purple), yielding the color purple. With more penicillin present, more PBP5 becomes inactive, and the purple gets lighter.