Team:HKU-Hong Kong/Project

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-<title>Introduction of our project 7-7-2011(amendments).docx</title><style type="text/css">@import url(https://themes.googleusercontent.com/fonts/css?kit=POVDFY-UUf0WFR9DIMCU8g);ol{margin:0;padding:0}p{margin:0}.c5{width:415.3pt;background-color:#ffffff;padding:72pt 90pt 72pt 90pt}.c0{font-size:12pt;font-style:italic;font-family:Droid Sans}.c1{text-align:justify;direction:ltr}.c3{font-size:12pt;font-family:Droid Sans}.c8{margin:5px;border:1px solid black}.c4{text-indent:36pt}.c6{height:12pt}.c7{font-weight:bold}.c2{direction:ltr}body{color:#000000;font-size:12pt;font-family:Times New Roman}h1{padding-top:12pt;line-height:1.0;text-align:left;color:#000000;font-size:16pt;font-family:Arial;font-weight:bold;padding-bottom:3pt}h2{padding-top:12pt;line-height:1.0;text-align:left;color:#000000;font-style:italic;font-size:14pt;font-family:Arial;font-weight:bold;padding-bottom:3pt}h3{padding-top:12pt;line-height:1.0;text-align:left;color:#000000;font-size:13pt;font-family:Arial;font-weight:bold;padding-bottom:3pt}h4{padding-top:12pt;line-height:1.0;text-align:left;color:#000000;font-size:14pt;font-family:Times New Roman;font-weight:bold;padding-bottom:3pt}h5{padding-top:12pt;line-height:1.0;text-align:left;color:#000000;font-style:italic;font-size:13pt;font-family:Times New Roman;font-weight:bold;padding-bottom:3pt}h6{padding-top:12pt;line-height:1.0;text-align:left;color:#000000;font-size:11pt;font-family:Times New Roman;font-weight:bold;padding-bottom:3pt}</style></head><body class="c5"><p class="c1"><span class="c3 c7">Introduction of our project</span></p><p class="c1 c6"><span class="c3 c7"></span></p><p class="c1"><span class="c3">&nbsp; &nbsp; Silencing of genes is a method adopted by cells to control their gene expression and the production of gene products. In eukaryotic cells, both heterochromatin and euchromatin are present. There is active transcription in euchromatin because of acetylation while there is inactive transcription in heterochromatin because of methylation. &nbsp; </span></p><p class="c1 c6"><span class="c3"></span></p><p class="c1"><span class="c3">&nbsp; &nbsp; Our project is called &ldquo;Super Silencer&rdquo;. We are trying to mimic heterochromatin in </span><span class="c0">Escherichia coli</span><span class="c3">&nbsp;(</span><span class="c0">E. coli</span><span class="c3">) so that we can better manipulate the bacteria by silencing particular genes. Our idea is to utilize</span><span class="c0">&nbsp;</span><span class="c3">a&nbsp;natural protein called histone-like nucleoid structuring protein (H-NS) to silence the target gene. H-NS is a major bacterial chromatin component which influences DNA structure and gene expression. It binds to DNA non-specifically with its C-terminal and polymerizes by its N-terminal.&nbsp;</span></p><p class="c1 c6"><span class="c3"></span></p><p class="c1 c4"><span class="c3">We are trying to replace&nbsp;the H-NS&#39;s&nbsp;C-terminal&nbsp;with a specific DNA binding domain like tetR to perform a site specific DNA binding to the corresponding DNA binding site tetO.&nbsp;We anticipate&nbsp;that the mutated H-NS with tetR will bind to the specific site (tetO) desired and recruit wild type H-NS (without tetR) to bind to adjacent DNA non-specifically and silence them on transcription level as a result of the blockage of gene by both the mutated and wild type H-SN. </span></p><p class="c1 c4 c6"><span class="c3"></span></p><p class="c1 c4"><span class="c3">We are using the green fluorescent protein (GFP) gene to test this principle. Normal genes without the binding of mutated H-SN with tetR to tetO site will have usual fluorescent intensity. However, if there is binding of mutated H-SN with tetR to the tetO site, there will be polymerisation of this specific DNA binding protein with the non-specific DNA binding protein. Hence, blocking of the transcription of the particular gene will result in silencing of the gene.
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