Team:Baskent-Meds

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                <strong>our</strong> research's pillars, <em> killing the legionella</em><a href="/Team:Baskent-Meds/Project">a link</a>.
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                <strong>Follows us </strong> in the <em>lab</em>, we are telling what we did <a href="/Team:Baskent-Meds/Notebook">a link</a>.
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                <strong>have</strong> we <em> met</em>? <a href="/Team:Baskent-Meds/Team">a link</a>.
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<li><a href="/Team:Baskent-Meds" title="BaskentMeds main page">Home</a></li>
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                <strong>also</strong> we have<em>fun!</em> with our competitors <a href="/Team:Baskent-Meds/Gallery">a link</a>.
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<a href="/Team:Baskent-Meds/Project" title="Our Project Abstract">Project</a>
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<li><a href="/Team:Baskent-Meds/Notebook" title="Our Beloved Notebook!">Notebook</a></li>
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<li><a href="/Team:Baskent-Meds/Results" title="Still Working on this">Results</a></li>
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<a href="/Team:Baskent-Meds/Team" title="Shall we meet?">Team</a>
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<li><a href="/Team:Baskent-Meds/Instructors" title="Our ">Instructors</a></li>
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  <p align="center" class="style10"><strong> Transformation of <em>Escherichia coli</em> In Order To Develop </strong></p>
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  <p align="center" class="style10"><strong><em> Legionella pneumophila</em> Sensing Bacteria </strong></p>
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  <p align="center"><strong> &nbsp;</strong></p>
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  <p> Our aim, as the team &ldquo;Baskent_Meds&rdquo;, is developing bacteria which can recognize <em>Legionella pneumophila</em> specifically at species level by legionella quorum sensing (lqs), and respond to that recognition by producing anti-Legionella peptide which is produced by some <em>Staphylococcus</em> strains. <em>lqs</em> gene locus is responsible for quorum sensing mechanism in<em> Legionella pneumophila</em>. On the <em>lqs</em> locus there are three genes; <em>lqsA</em> (encoding autoinducer synthase), <em>lqsR</em> (encoding response regulator), and <em>lqsS</em> (encoding a sensor kinase). LqsA is a pyridoxal-5&prime;-phosphate-dependent enzyme that catalyses the production of the signaling molecule 3-hydroxypentadecane-4-one (LAI-1; Legionella autoinduer-1), a novel member of &alpha;-hydroxyketone signalling molecules (Spirig et al., 2008). In this comprehensive and long-termed project, our initial objectives were; generating competent <em>E. coli</em> cells in which we are planning to clone <em>lqs</em> gene locus (the gene locus responsible for quorum sensing mechanism in<em> Legionella pneumophila</em>), and optimizing transformation procedures. pNT-1 (Tiaden et al., 2007), a pUCBM20-derivative, bearing full <em>lqs</em> gene cluster, and pTS-2 (Spirig et al., 2008), a pMMB207C-derivative expression vector, bearing <em>LqsA</em> gene were kindly provided by Prof. Dr. Hubert Hilbi. </p>
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  <p> Initially, <em>E. coli</em> competent cell groups (JM109, DH5&alpha; and XL1-Blue) were formed by exerting calcium chloride precipitation. pNT-1 and pTS-2 were transferred into competent <em>E. coli</em> cells using heat shock transformation method, and clones were selected with ampicillin and chloramphenicol, respectively. Plasmid isolations were performed from transformants by QIAprep Spin Miniprep Kit (Qiagen), and visualized by agarose gel electrophoresis. Our recent studies are designing our construct with BioBricks, and amplifying <em>LqsR</em> and <em>LqsS</em> by gene specific primers having restriction enzyme cut sites as small adaptors. We are going to clone these genes by digesting with these restriction enzymes followed by ligation to one of the iGEM vectors. After cloning these genes, we are planning to clone anti-Legionella peptide nucleotide sequence under <em>lqs</em> control. In order to check response of transformant <em>E. coli</em> we are planning to clone pTS-2 to BL21(DE3) <em>E. coli</em> for the constitutive expression and production of the LAI-1 without the handling of the pathogen strains in our laboratory. </p>
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  <p> By the end of our experiments, we aim to obtain sensor <em>E. coli</em> cells which respond to <em>Legionella pneumophila </em>quorum sensing by recognizing LAI-1. Our expression construct will also enable production of the anti-Legionella peptide in response to LAI-1. Since the peptide has specific antibacterial activity on <em>Legionella</em> spp., the achievement of the project is the destruction of <em>Legionella pneumophila </em>by transformant <em>E. coli</em> sensor cells. </p>
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Revision as of 01:45, 1 August 2012

Smooth Animated jQuery Menu

our research's pillars, killing the legionellaa link.
Follows us in the lab, we are telling what we did a link.
have we met? a link.
also we havefun! with our competitors a link.