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| | <h3>Before you start</h3> | | <h3>Before you start</h3> |
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| | <li> <a href="https://2018.igem.org/Resources/Template_Documentation">Template documentation</a></li> | | <li> <a href="https://2018.igem.org/Resources/Template_Documentation">Template documentation</a></li> |
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| − | <h3> Construct the promoter library </h3>
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| − | <p>Our team will use Pseudomonas Fluorescens Pf-5 as a chassis (Pseudomonas Fluorescens Pf-5 is one of the microorganisms that can be obtained commercially by the Chinese Ministry of Agriculture), and construct the promoter library consists of strong, medium, and weak promoters. Not only will it ensure the normal expression of heterologous genes in Pseudomonas fluorescens, but also can regulate the expression intensity of the heterologous genes, and provide a basis for the construction of standardized Pseudomonas fluorescens vectors.</p>
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| − | <h3> Treatment of cigarette butts with degrading bacteria </h3>
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| − | <p>In the 1950s, smoking caused a large increase in cancer patients. Tobacco companies developed filters(They can effectively reduce the wastes as tar and nicotine in flue gas. ) It is estimated that the number of filters consumed each year in the world reaches 5.6 trillion, and China has exceeded 180 million. some experts even recommend that cigarette butts be managed as hazardous wastes.</p>
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| − | <p>Pseudomonas Fluorescens Pf-5 itself has been able to degrade tar, phenols, organic chlorine and other contaminants. In our project, a large fragment was directly cloned using the Red/ET recombination system, and the nicotine degradation gene cluster of Agrobactrium tumefaciens S33 was introduced into Pseudomonas Fluorescens Pf-5 to increase the nicotine degradation ability.We strongly believe that our project has great application value in the centralized treatment of cigarette butts and the restoration of cigarette butt-polluted environment.</p>
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| − | <h3> Combination and Application </h3>
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| − | <p>SKLMT-China is going to transform P. fluorescens to provide a series of standardized promoter elements with different strength,in tihs way to regulate gene expression in P.fluorescens this year. At the same time, we will use the Red/ET recombination system(It can direct the cloning of large fragments) to introduce the nicotine degradation gene cluster in Agrobactrium tumefaciens S33 into Pseudomonas Fluorescens Pf-5, and achieve the heterologous expression by using the promoter element we are going to construct. Finally, the cigarette pollution degrading bacteria which can be directly released into the environment will be structured.</p>
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