Difference between revisions of "Team:Hangzhou-H14Z"
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+ | <li><a href="#tf-home" class="scroll">Home</a></li> | ||
+ | <li><a href="#tf-intro" class="scroll">intro</a></li> | ||
+ | <li><a href="#tf-services" class="scroll">PROJECT</a></li> | ||
+ | <li><a href="#tf-works" class="scroll">TEAM</a></li > | ||
+ | <li><a href="#tf-process" class="scroll">policy</a></li> | ||
+ | <li><a href="#tf-pricing" class="scroll">support</a></li> | ||
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− | < | + | <h1 >The best way ever to preserve lithic relics </h1> |
− | + | <p class="lead">an innovative biochemical method to resolve erosion. </p> | |
− | < | + | <a href="#tf-intro" class="scroll goto-btn text-uppercase">View Our Works</a> <!-- Link to your portfolio section --> |
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+ | <div class="row"> <!-- row --> | ||
− | < | + | <div class=" col-md-offset-2 col-lg-offset-0 col-lg-11"> |
+ | <br> | ||
+ | <br> | ||
+ | <img src="img/logo-w.png" class="intro-logo img-responsive" alt="free-template"> <!-- Your company logo in white --> | ||
+ | <p>The inscriptions on Precipice are one of the most common and fundamental forms of traditional Chinese relics. Due to having been carved in open areas plus the exposure to sunshine and raindrops, which is often the case for most of them,the Inscriptions are weathered readily. Currently, there are quite a few methods to resolve this phenomenon but some of them are not used properly which occasionally cause the damage for the Inscriptions. Our research project aims at using microorganism method, which is to put a plasmid with the ability to convert bio-oriented oxaloacetic acid to oxalate acid into the cell of E.coil, inducing the cell and the product will react with one component of the Inscriptions-- calcium carbonate-- to generate nearly insoluble calcium oxalate monohydrate to adhere to the Inscriptions for protection. Another E.coil with the ablility to secrete lipase has also been constructed; we aim to add this cell when we are cleansing the Inscriptions to resolve the oil substances on the surface of them. By constructing these products, we are able to protect the relics either from human errors as cleaning and protecting or from natural damnification. </p> | ||
+ | </div> | ||
+ | |||
+ | </div><!-- end row --> | ||
+ | </div> | ||
+ | <div class="container"> <!-- container --> | ||
+ | <div class="section-header"> | ||
+ | <h2>some   <span class="highlight"><strong><font color="#00C459">destroyed   </font></strong></span> inscriptions   on   Precipice</h2> | ||
+ | <h5><em>photos were took within Hangzhou</em></h5> | ||
+ | <div class="fancy"><span><img src="img/favicon.ico" alt="..."></span></div> | ||
+ | </div> | ||
+ | <div id="team" class="owl-carousel owl-theme text-center"> <!-- team carousel wrapper --> | ||
+ | <div class="item"><!-- Team #1 --> | ||
+ | <div class="hover-bg"> <!-- Team Wrapper --> | ||
+ | |||
+ | <img src="img/DSC_0172.jpg" alt="..." class="img-responsive"> <!-- Team Image --> | ||
+ | <div class="team-detail text-center"> | ||
+ | <h3>The White Tower</h3> | ||
+ | <p class="text-uppercase">a famous religious symbol in Hangzhou</p> | ||
+ | |||
+ | </div> | ||
+ | </div><!-- End Team Wrapper --> | ||
+ | </div><!-- End Team #1 --> | ||
− | < | + | <div class="item"> <!-- Team #2 --> |
+ | <div class="hover-bg"> <!-- Team Wrapper --> | ||
+ | |||
+ | <img src="img/DSC_0086.jpg" alt="..." class="img-responsive"><!-- Team Image --> | ||
+ | <div class="team-detail text-center"> | ||
+ | <h3>Figure of Buddha</h3> | ||
+ | <p class="text-uppercase">a symbol of Buddhism</p> | ||
+ | |||
+ | </div> | ||
+ | </div> <!-- End Team Wrapper --> | ||
+ | </div><!-- End Team #2 --> | ||
+ | <div class="item"> <!-- Team #3 --> | ||
+ | <div class="hover-bg"> <!-- Team Wrapper --> | ||
+ | |||
+ | <img src="img/DSC_0150.jpg" alt="..." class="img-responsive"><!-- Team Image --> | ||
+ | <div class="team-detail text-center"> | ||
+ | <h3>Woman in palace</h3> | ||
+ | <p class="text-uppercase">stone inscription</p> | ||
+ | |||
+ | </div> | ||
+ | </div><!-- End Team Wrapper --> | ||
+ | </div><!-- End Team #3 --> | ||
+ | |||
+ | <div class="item"><!-- Team #4 --> | ||
+ | <div class="hover-bg"> <!-- Team Wrapper --> | ||
+ | |||
+ | <img src="img/DSC_0213.jpg" alt="..." class="img-responsive"> <!-- Team Image --> | ||
+ | <div class="team-detail text-center"> | ||
+ | <h3>Statue of MAHAKALA</h3> | ||
+ | <p class="text-uppercase">Ancient monument</p> | ||
+ | |||
+ | </div> | ||
+ | </div> <!-- End Team Wrapper --> | ||
+ | </div><!-- End Team #4 --> | ||
+ | |||
+ | <div class="item"><!-- Team #5 --> | ||
+ | <div class="hover-bg"> <!-- Team Wrapper --> | ||
+ | |||
+ | <img src="img/DSC_0224.jpg" alt="..." class="img-responsive"> <!-- Team Image --> | ||
+ | <div class="team-detail text-center"> | ||
+ | <h3>Ancient chinese governor</h3> | ||
+ | <p class="text-uppercase">stone inscription</p> | ||
+ | |||
+ | </div> | ||
+ | </div> <!-- End Team Wrapper --> | ||
+ | </div><!-- End Team #5 --> | ||
+ | |||
+ | </div> <!-- end team carousel wrapper --> | ||
+ | |||
+ | </div> <!-- container --><!-- end container --> | ||
+ | </div> | ||
+ | |||
+ | <!-- Service Section | ||
+ | ================================================== --> | ||
+ | <div id="tf-services"> | ||
+ | <div class="container"> <!-- container --> | ||
+ | |||
+ | <div class="section-header"> | ||
+ | <h2>Project's<span class="highlight"><strong> Presentation</strong></span></h2> | ||
+ | <h5><em>photoes were took within Hangzhou</em></h5> | ||
+ | <div class="fancy"><span><img src="img/favicon.ico" alt="..."></span></div> | ||
+ | |||
+ | |||
+ | </div><!-- end container --> | ||
+ | </div> | ||
+ | |||
+ | <!-- About Us Section | ||
+ | ================================================== --> | ||
+ | |||
+ | |||
+ | <div id="feature" class="gray-bg"> <!-- fullwidth gray background --> | ||
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+ | <div class="row" role="tabpanel"> <!-- row --> | ||
+ | <div class="col-md-4 col-md-offset-1"> <!-- tab menu col 4 --> | ||
+ | |||
+ | <ul class="features nav nav-pills nav-stacked" role="tablist"> | ||
+ | <li role="presentation" class="active"> <!-- feature tab menu #1 --> | ||
+ | <a href="#f1" aria-controls="f1" role="tab" data-toggle="tab"> | ||
+ | Project's background part I<br> | ||
+ | </a> | ||
+ | </li> | ||
+ | <li role="presentation"> <!-- feature tab menu #3 --> | ||
+ | <a href="#f3" aria-controls="f3" role="tab" data-toggle="tab"> | ||
+ | Project's background part II<br> | ||
+ | </a> | ||
+ | </li> | ||
+ | <li role="presentation"> <!-- feature tab menu #4 --> | ||
+ | <a href="#f4" aria-controls="f4" role="tab" data-toggle="tab"> | ||
+ | Principles of experiments<br> | ||
+ | </a> | ||
+ | </li> | ||
+ | <li role="presentation"> <!-- feature tab menu #5 --> | ||
+ | <a href="#f5" aria-controls="f5" role="tab" data-toggle="tab"> | ||
+ | Procedures in detail<br> | ||
+ | </a> | ||
+ | </li> | ||
+ | <li role="presentation" > <!-- feature tab menu #1 --> | ||
+ | <a href="#f6" aria-controls="f6" role="tab" data-toggle="tab"> | ||
+ | Mathimatical model<br> | ||
+ | </a> | ||
+ | </li> | ||
+ | <li role="presentation" > <!-- feature tab menu #1 --> | ||
+ | <a href="#f7" aria-controls="f7" role="tab" data-toggle="tab"> | ||
+ | Time line<br> | ||
+ | </a> | ||
+ | </li> | ||
+ | </ul> | ||
+ | |||
+ | </div><!-- end tab menu col 4 --> | ||
+ | |||
+ | <div class="col-md-6"> <!-- right content col 6 --> | ||
+ | <!-- Tab panes --> | ||
+ | <div class="tab-content features-content"> <!-- tab content wrapper --> | ||
+ | <div role="tabpanel" class="tab-pane fade in active" id="f1"> <!-- feature #1 content open --> | ||
+ | <h4>Project's background part I</h4> | ||
+ | <p> The area near Qiantang River has a profound history of wealth. Hangzhou, in the heart of this area, situated many astonishing lithic cultural relics. Most of them were made during The Ten Kingdoms (A.D. 891-A.D. 979) and Song Dynasty (A.D. 960- A.D. 1279). Not only do those lithic relics represent the cutting edge of stone carving technology in contemporary time, but also they provide historians and archaeologists with solid historical evidence. </p><p> | ||
+ | There are three main types of lithic relics in Hangzhou; they are stone sculptures, stone towers also known as Dhavaja, and stone inscriptions. The stone sculptures are primarily represented by Feilai Peak (飞来峰), Ciyun Ridge (慈云岭), and Statue of Mahakala in Baocheng Temple (宝成寺麻曷葛剌). Stone Towers are primarily represented by White Tower (白塔). Stone inscriptions are primarily represented by Hetingwuhe Fubei on Lonely Mountain (孤山放鹤亭舞鹤赋碑).</p><p> Plus, Feilai Peak and White Tower are National Key Historical Relics. | ||
+ | Most lithic relics encounter physical, chemical and biological interaction, resulting in weathering corrosion, pulverisation lesion, cracking, contamination, and colour mottling. Those phenomena jeopardise the value of lithic relics and even lithic relics themselves, and the lithic relics in Hangzhou are not of exceptions. <p/><p> | ||
+ | In 2002, some of the classic lithic relics were inspected and it was found that the surface of Feilai Peak Sculpture was greatly ruined (Figure 1), and there is leaking on Ciyun Ridge Sculpture (Figure 2)..</p> <p> Meanwhile, Cultural Relics Department of China has been trying to maintain and recover the lithic relics. Since 2002, Cultural Relics Department of China mainly has been removing surface deposit, depleting pounding, strengthening crag, and changing flora; the government washed Feilai Peak Sculpture for many times (Figure 3).Government built a top above the Ciyun Peak Sculpture, this relic no longer had to expose to sun and rain henceforward (Figure 4).</p><p> White Tower was comprehensively cleaned and strengthened by silicone by Zhejiang University of Technology (Figure 5).</p> | ||
+ | <div id="myCarousel" class="carousel slide"> | ||
+ | <ol class="carousel-indicators"> | ||
+ | <li data-target="#myCarousel" data-slide-to="0" class="active"></li> | ||
+ | <li data-target="#myCarousel" data-slide-to="1"></li> | ||
+ | <li data-target="#myCarousel" data-slide-to="2"></li> | ||
+ | <li data-target="#myCarousel" data-slide-to="3"></li> | ||
+ | <li data-target="#myCarousel" data-slide-to="4"></li> | ||
+ | <li data-target="#myCarousel" data-slide-to="5"></li> | ||
+ | <li data-target="#myCarousel" data-slide-to="6"></li> | ||
+ | <li data-target="#myCarousel" data-slide-to="7"></li> | ||
+ | </ol> | ||
+ | <div class="carousel-inner"> | ||
+ | <div class="item active"> | ||
+ | <img src="img/relics/fg1.png" alt="First slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 1</p></center> | ||
+ | </div> | ||
+ | |||
+ | <div class="item"> | ||
+ | <img src="img/relics/fg2.png" alt="Second slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 2</p></center> | ||
+ | </div> | ||
+ | <div class="item"> | ||
+ | <img src="img/relics/fg31.png" alt="First slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 3-1</p></center> | ||
+ | </div> | ||
+ | |||
+ | <div class="item"> | ||
+ | <img src="img/relics/fg32.png" alt="Second slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 3-2</p></center> | ||
+ | </div> | ||
+ | <div class="item"> | ||
+ | <img src="img/relics/fg41.png" alt="Second slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 4-1</p></center> | ||
+ | </div> | ||
+ | <div class="item"> | ||
+ | <img src="img/relics/fg42.png" alt="Second slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 4-2</p></center> | ||
+ | </div> | ||
+ | <div class="item"> | ||
+ | <img src="img/relics/fg51.png" alt="Second slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 5-1</p></center> | ||
+ | </div> | ||
+ | <div class="item"> | ||
+ | <img src="img/relics/fg52.png" alt="Second slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 5-2</p></center> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div role="tabpanel" class="tab-pane fade" id="f3"> <!-- feature #2 content --> | ||
+ | <h4>Project's background part II</h4> | ||
+ | <p> The power plants and railways were removed. During the washing, the staff intentionally left part of this tower intact, and made an contrast for further research of anti-contamination and anti-weathering (Figure 5-3, 5-4)</p><p> However, 2015 field research shows most external lithic murals, statue, and towers still suffer from damage. Some even have distraction in main stone body. Meanwhile, there grows moss. Indoor stone inscriptions though are visible; there are obvious and irremovable cracks. </p><p> | ||
+ | Today, most preservation of lithic relics employ physical, chemical, or engineer approaches no matter where. It has been proven by reality that each of those approaches has fatal disadvantages for long-standing preservation of lithic relics, and those disadvantages would even lead to the ruin of lithic relics.</p> <p> Since forever, there are two categories of methods to preserve-- organic and inorganic. Organic materials cannot reach the standard of time to preserve relics such as epoxy resin, acrylic resin, silicone, etc.; inorganic materials would form hard shell that is not compatible or even react with the lithic relics, such as lime, barium hydroxide, and sulphuric acid. </p><p> | ||
+ | In conclusion, finding a long-standing, inexpensive, and efficient method to preserve lithic relics is an impending issue. </p> | ||
+ | <div id="myCarouselt" class="carousel slide"> | ||
+ | <ol class="carousel-indicators"> | ||
+ | <li data-target="#myCarouselt" data-slide-to="0" class="active"></li> | ||
+ | <li data-target="#myCarouselt" data-slide-to="1"></li> | ||
+ | </ol> | ||
+ | <div class="carousel-inner"> | ||
+ | <div class="item active"> | ||
+ | <img src="img/relics/fg53.png" alt="First slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 5-3</p></center> | ||
+ | </div> | ||
+ | |||
+ | <div class="item"> | ||
+ | <img src="img/relics/fg54.png" alt="Second slide"> | ||
+ | <br> | ||
+ | <center><p>Figure 5-4</p></center> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div role="tabpanel" class="tab-pane fade" id="f4"> <!-- feature #2 content --> | ||
+ | <h4>Principles of experiments</h4> | ||
+ | <p> The documents cited shows that even after natural weathering, biological decomposition, and acid rain erosion for a time period of time, some stone inscriptions, which are on rock surface containing calcium, remain in good condition because there is a thin layer of naturally formed semitransparent mineralised film that is hydrophilic and compact on such rock surface.</p><p> In some part under the fine film, knife scratches on the stone from over a thousand years ago are still visible indistinctly. According to solid analysis, natural mineralised film is primarily composed of calcium oxalate monohydrate [Ca(COO)2·H2O] whose process of composition is interfered by microorganism.</p><p> | ||
+ | There are some advantages of calcium oxalate being a film as follows,</p> | ||
+ | <p> 1) In water, especially in acidulous water, the solubility of calcium oxalate is one order of magnitude smaller than normal calcium carbonate.</p> | ||
+ | <p> 2) Organically oriented calcium oxalate crystal's particles are fine and compact, which prevent the infiltration of toxic and harmful materials. </p> | ||
+ | <p> 3) The system can't be easily broken by external forces in that there is no distinct boundary between calcium carbonate rock and calcium oxalate film.</p> | ||
+ | <p> So, this research project has chosen E. coli that has been transcribed with lipase to clean some stone relics that are covered with oil; then, we use E. coli, with transcription, that can secrete oxalate acid upon the stone relics. Therefore, there forms calcium oxalate to be protection film in order to preserve lithic relics.</p> | ||
+ | <p> If we use chemical methods to generate calcium oxalate, we have to immerse the lithic relics in the chemical liquid; however, it is not suitable for the preservation of large-scale outdoor lithic relics. In fact, the natural existing calcium oxalate comes from the reaction of oxalate acid generated by microorganism and calcium compound on the surface of stone. This research project will utilise genetic engineering methods to "graft" the gene that generates calcium oxalate into Escherichia coli (E. coil).</p><p> Then oxalate acid reacts with calcium lithic relics to generate calcium oxalate, which stimulates its natural formation process. Finally, there forms a fine layer to protect the lithic relics. </p> | ||
+ | </div> | ||
+ | <div role="tabpanel" class="tab-pane fade" id="f5"> <!-- feature #2 content --> | ||
+ | <h4>Procedures in details</h4> | ||
+ | <p> | ||
+ | 4 实验过程</p> | ||
+ | <p> 4.1 实验试剂及仪器</p> | ||
+ | <p> 4.1.1 目的基因 草酰乙酸乙酰基水解酶(oxaloacetate acetyl hydrolase 简写OAH)是本实验所使用的目的基因片段,主要功效是在表达时将草酰乙酸转化为草酸,草酸进而于钙结合生成草酸钙起到文物保护的作用。此目的基因片段从基因库中查的基因序列后由美国IDT公司直接合成得到。 </p> | ||
+ | <p> 4.1.2 菌种和质粒 本实验使用的质粒为pET-28a,使用的大肠杆菌为TOP-10感受态细胞,购于天根生化科技有限公司</p> | ||
+ | <p> 4.1.3 酶与试剂 实验所使用的限制性内切酶购于NEB,连接酶购于天根生化,DNA纯化试剂盒以及质粒提取试剂盒均购于天根生化。</p> | ||
+ | <p> 4.1.4 PCR引物 根据OAH以测得的序列,由Invitrogen合成了如下引物:</p> | ||
+ | <p> 上游引物:5’-ATCATCTTGCATATGGCGTCCACCATTGCTGTT-3’(划线部分为Nde i酶切位点)</p> | ||
+ | <p> 下游引物:5’-TTTCCTTTTGCGGCCGCTTAG-3’(划线部分为Not i酶切位点)</p> | ||
+ | <p> 4.1.5 培养基 实验使用的培养基为LB,以配备200ml的量为例,需要如下材料:胰蛋白胨2g,酵母提取物1g,NaCl 2g,加水至200ml。</p> | ||
+ | <p> 实验使用的培养板加入卡那抗性,一配备400ml为例,需要以下材料:胰蛋白胨 4g,酵母提取物 2g,NaCl 4g,Agar琼脂粉6g,卡那霉素 1.2ml</p> | ||
+ | <p> 4.2 实验步骤</p> | ||
+ | <p> 4.2.1 PCR扩增 因为目的基因片段的G-C碱基对含量很高,所以PCR扩增时需使用GC Buffer,参照Takara产品目录配200ul体系:GC buffer 100ul,dntp mixture 16ul, 前引物 4ul, 后引物 4ul,DNA模板 4ul, Taq 2ul, H2O 70ul。用60℃-70℃进行梯度扩增,寻找引物合适的退火温度。发现在第六条带最亮,再按照上述配成新的体系在适宜温度下进行PCR扩增,得到加上接头的基因片段</p> | ||
+ | <p> 4.2.2 做完PCR之后,因有较多的杂条带,故将所有扩增得到的产物跑胶,将所需片段切胶纯化。</p> | ||
+ | <p> 4.2.3 进行酶切,分别在目的基因片段和载体pET-28a中加入Not I 和 Nde I 的限制性内切酶。</p> | ||
+ | <p> 4.2.4 进行连接并设置对照组。实验组:大片段+小片段+T4 DNA Ligase+Buffer</p> | ||
+ | <p> 对照组:大片段 | ||
+ | 在16℃的环境下连接过夜。</p> | ||
+ | <p> 4.2.5 使用DNA纯化试剂盒提取连接后产物,转化进TOP-10感受态细胞,涂在配置好的带卡那抗性的培养板上,放在37℃的恒温环境下培养。</p> | ||
+ | <p> 4.2.6 根据菌落的生长情况,将实验组的细菌进行质粒提取,并再次用上述的两种酶进行酶切后跑胶,如果跑胶后大片段和小片段的位置正确,则连接完成。</p> | ||
+ | <p> 4.2.7 测序。</p> | ||
+ | <p> 4.3 实验结果 | ||
+ | </p> | ||
+ | </div> | ||
+ | |||
+ | </div> <!-- end tab content wrapper --> | ||
+ | </div><!-- end right content col 6 --> | ||
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+ | |||
+ | <!-- Works Section | ||
+ | ================================================== --> | ||
+ | <div id="tf-works"> | ||
+ | <div class="container"> | ||
+ | <div class="section-header"> | ||
+ | <h2>iNFO ABOUT <span class="highlight"><strong>Team MEMBER</strong></span></h2> | ||
+ | <h5><em>suspend your mouse above each images to learn more about our team member.</em></h5> | ||
+ | <div class="fancy"><span><img src="img/favicon.ico" alt="..."></span></div> | ||
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+ | |||
+ | <div class="text-center"> | ||
+ | <ul class="list-inline cat"> <!-- Portfolio Filter Categories --> | ||
+ | <li><a href="#" data-filter="*" class="active">Team members</a></li> | ||
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+ | </ul><!-- End Portfolio Filter Categories --> | ||
+ | </div> | ||
+ | |||
+ | </div><!-- End Container --> | ||
+ | |||
+ | <div class="container-fluid"> <!-- fluid container --> | ||
+ | <div id="itemsWork" class="row text-center"> <!-- Portfolio Wrapper Row --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 col-lg-3 nopadding brand others"> <!-- Works #1 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> <!-- Hover Description --> | ||
+ | |||
+ | <p><h10>Zhengdong Chen</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/dongdong.png" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/dongdong.png"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div><!-- End Hover Description --> | ||
+ | <img src="img/portfolio/01.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div><!-- end Works #1 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 col-lg-3 nopadding apps"> <!-- Works #2 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p><h10>Yue Wu</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/wuyue.png" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/wuyue.png"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | </div> | ||
+ | <img src="img/portfolio/02.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div><!-- end Works #2 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 col-lg-3 nopadding others brand"><!-- Works #3 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p><h10>Shuyang Zhou</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/shuyangzhou.png" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/shuyangzhou.png"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/03.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div><!-- end Works #3 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 col-lg-3 nopadding others web"> <!-- Works #4 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p><h10>Haobo Shao</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/haoboshao.png" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/haoboshao.png"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/04.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> <!-- end Works #4 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 nopadding web others"> <!-- Works #5 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p><h10>Yunfeng Guo</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/05@2x.jpg" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/05@2x.jpg"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/05.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> <!-- end Works #5 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 nopadding app"> <!-- Works #6 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p><h10>Minxing Chen</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/minxingchen.png" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/minxingchen.png"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/06.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div><!-- end Works #6 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 nopadding web brand"><!-- Works #7 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p> <h10>Jingcheng Shi</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/jingchengshi.png" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/0jingchengshi.png"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/07.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div><!-- end Works #7 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 nopadding app"> <!-- Works #8 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p><h10>Juncheng Luo</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/junchengluo.png" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/junchengluo.png"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/08.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> <!-- end Works #8 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 nopadding web brand"><!-- Works #7 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p> <h10>Wei He</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/weihe.png" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/weihe.png"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/10.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div><!-- end Works #7 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 nopadding web brand"><!-- Works #7 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p> <h10>Yuning Gu</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/07@2x.jpg" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/07@2x.jpg"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/11.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div><!-- end Works #7 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 nopadding web brand"><!-- Works #7 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p> <h10>Xiaomei Tang</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/07@2x.jpg" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/07@2x.jpg"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/07.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div><!-- end Works #7 col 3 --> | ||
+ | |||
+ | <div class="col-xs-12 col-sm-6 col-md-3 nopadding web brand"><!-- Works #7 col 3 --> | ||
+ | <div class="item"> | ||
+ | <div class="hover-bg"> | ||
+ | <div class="hover-text off"> | ||
+ | <p> <h10>Jie Zheng</h10></p> | ||
+ | <a title="Freedom Project #1" href="img/portfolio/07@2x.jpg" data-lightbox-gallery="gallery1" data-lightbox-hidpi="img/portfolio/07@2x.jpg"> | ||
+ | <i class="fa fa-expand"></i> | ||
+ | </a> | ||
+ | |||
+ | </div> | ||
+ | <img src="img/portfolio/07.jpg" class="img-responsive" alt="Image"> <!-- Portfolio Image --> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div><!-- end Works #7 col 3 --> | ||
+ | |||
+ | |||
+ | </div> <!-- End Row --> | ||
+ | |||
+ | </div> <!-- End Container-Fluid --> | ||
+ | </div> | ||
+ | |||
+ | <!-- Process Section | ||
+ | ================================================== --> | ||
+ | <div id="tf-process"> | ||
+ | <div class="container"> <!-- container --> | ||
+ | <div class="section-header"> | ||
+ | <h2>Policy & <span class="highlight"><strong>Human practices</strong></span></h2> | ||
+ | <h5><em>We design and build functional and beautiful websites</em></h5> | ||
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+ | ================================================== --> | ||
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+ | <h2>The <span class="highlight"><strong>Support </strong></span>of our team and project</h2> | ||
+ | <h5><em>This investigative project is supported by various well-known institutes, universities and corporations</em></h5> | ||
+ | <div class="fancy"><span><img src="img/favicon.ico" alt="..."></span></div> | ||
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+ | ================================================== --> | ||
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+ | <div class="container"> <!-- container --> | ||
+ | <div class="section-header"> | ||
+ | <h2>The impact we made about <span class="highlight"><strong> synthetic biology</strong></span></h2> | ||
+ | <h5><em>We build a set of pages in our high school's official website to publicize the subject of synthetic biology</em></h5> | ||
+ | <div class="fancy"><span><img src="img/favicon.ico" alt="..."></span></div> | ||
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+ | <div class="media-right media-middle"> | ||
+ | <a href="http://ap.h14z.com/igem/index.html"> <i class="fa fa-hand-o-right"></i></a> | ||
+ | </div> | ||
+ | <div class="media-body"> | ||
+ | <a href="http://ap.h14z.com/igem/index.html"><h4 class="media-heading">The page of H14z's synthetic biology</h4></a> | ||
+ | <a href="http://ap.h14z.com/igem/index.html"><p>This page was built by Minxing Chen as well. There is a brief introduction about the synthetic biology and it was written in simplified chinese. Also, we put some useful knowledges there such as the bio-switch. Abundant people has visited this website and learnd a lot about synthetic biology and our school.</p></a> | ||
+ | </div> | ||
+ | </div><!-- Process #1 --> | ||
+ | |||
+ | <div class="media process"> <!-- Process #1 --> | ||
+ | <div class="media-right media-middle"> | ||
+ | <a href="http://ap.h14z.com"> <i class="fa fa-hand-o-right"></i></a> | ||
+ | </div> | ||
+ | <div class="media-body"> | ||
+ | <a href="http://ap.h14z.com"><h4 class="media-heading">AP center's website</h4></a> | ||
+ | <a href="http://ap.h14z.com"><p>All of our team members came from Hangzhou #14 highschool's Ap center. And we also put the synthetic biology's link there. Consequently, more students and facilities can see it.</p></a> | ||
+ | </div> | ||
+ | </div><!-- Process #1 --> | ||
+ | <div class="media process"> <!-- Process #1 --> | ||
+ | <div class="media-right media-middle"> | ||
+ | <a href="http://www.h14z.com"> <i class="fa fa-hand-o-right"></i></a> | ||
+ | </div> | ||
+ | <div class="media-body"> | ||
+ | <a href="http://www.h14z.com"><h4 class="media-heading">The official website of Hangzhou #14 high school</h4></a> | ||
+ | <a href="http://www.h14z.com"><p>This is the offcial website of our school. And you can find aboved links there.</p></a> | ||
+ | </div> | ||
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Revision as of 05:21, 15 September 2015
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The inscriptions on Precipice are one of the most common and fundamental forms of traditional Chinese relics. Due to having been carved in open areas plus the exposure to sunshine and raindrops, which is often the case for most of them,the Inscriptions are weathered readily. Currently, there are quite a few methods to resolve this phenomenon but some of them are not used properly which occasionally cause the damage for the Inscriptions. Our research project aims at using microorganism method, which is to put a plasmid with the ability to convert bio-oriented oxaloacetic acid to oxalate acid into the cell of E.coil, inducing the cell and the product will react with one component of the Inscriptions-- calcium carbonate-- to generate nearly insoluble calcium oxalate monohydrate to adhere to the Inscriptions for protection. Another E.coil with the ablility to secrete lipase has also been constructed; we aim to add this cell when we are cleansing the Inscriptions to resolve the oil substances on the surface of them. By constructing these products, we are able to protect the relics either from human errors as cleaning and protecting or from natural damnification.
some   destroyed   inscriptions   on   Precipice
photos were took within Hangzhou
![...](img/DSC_0172.jpg)
The White Tower
a famous religious symbol in Hangzhou
![...](img/DSC_0086.jpg)
Figure of Buddha
a symbol of Buddhism
![...](img/DSC_0150.jpg)
Woman in palace
stone inscription
![...](img/DSC_0213.jpg)
Statue of MAHAKALA
Ancient monument
![...](img/DSC_0224.jpg)
Ancient chinese governor
stone inscription
Project's Presentation
photoes were took within Hangzhou
Project's background part I
The area near Qiantang River has a profound history of wealth. Hangzhou, in the heart of this area, situated many astonishing lithic cultural relics. Most of them were made during The Ten Kingdoms (A.D. 891-A.D. 979) and Song Dynasty (A.D. 960- A.D. 1279). Not only do those lithic relics represent the cutting edge of stone carving technology in contemporary time, but also they provide historians and archaeologists with solid historical evidence.
There are three main types of lithic relics in Hangzhou; they are stone sculptures, stone towers also known as Dhavaja, and stone inscriptions. The stone sculptures are primarily represented by Feilai Peak (飞来峰), Ciyun Ridge (慈云岭), and Statue of Mahakala in Baocheng Temple (宝成寺麻曷葛剌). Stone Towers are primarily represented by White Tower (白塔). Stone inscriptions are primarily represented by Hetingwuhe Fubei on Lonely Mountain (孤山放鹤亭舞鹤赋碑).
Plus, Feilai Peak and White Tower are National Key Historical Relics. Most lithic relics encounter physical, chemical and biological interaction, resulting in weathering corrosion, pulverisation lesion, cracking, contamination, and colour mottling. Those phenomena jeopardise the value of lithic relics and even lithic relics themselves, and the lithic relics in Hangzhou are not of exceptions.
In 2002, some of the classic lithic relics were inspected and it was found that the surface of Feilai Peak Sculpture was greatly ruined (Figure 1), and there is leaking on Ciyun Ridge Sculpture (Figure 2)..
Meanwhile, Cultural Relics Department of China has been trying to maintain and recover the lithic relics. Since 2002, Cultural Relics Department of China mainly has been removing surface deposit, depleting pounding, strengthening crag, and changing flora; the government washed Feilai Peak Sculpture for many times (Figure 3).Government built a top above the Ciyun Peak Sculpture, this relic no longer had to expose to sun and rain henceforward (Figure 4).
White Tower was comprehensively cleaned and strengthened by silicone by Zhejiang University of Technology (Figure 5).
Project's background part II
The power plants and railways were removed. During the washing, the staff intentionally left part of this tower intact, and made an contrast for further research of anti-contamination and anti-weathering (Figure 5-3, 5-4)
However, 2015 field research shows most external lithic murals, statue, and towers still suffer from damage. Some even have distraction in main stone body. Meanwhile, there grows moss. Indoor stone inscriptions though are visible; there are obvious and irremovable cracks.
Today, most preservation of lithic relics employ physical, chemical, or engineer approaches no matter where. It has been proven by reality that each of those approaches has fatal disadvantages for long-standing preservation of lithic relics, and those disadvantages would even lead to the ruin of lithic relics.
Since forever, there are two categories of methods to preserve-- organic and inorganic. Organic materials cannot reach the standard of time to preserve relics such as epoxy resin, acrylic resin, silicone, etc.; inorganic materials would form hard shell that is not compatible or even react with the lithic relics, such as lime, barium hydroxide, and sulphuric acid.
In conclusion, finding a long-standing, inexpensive, and efficient method to preserve lithic relics is an impending issue.
Principles of experiments
The documents cited shows that even after natural weathering, biological decomposition, and acid rain erosion for a time period of time, some stone inscriptions, which are on rock surface containing calcium, remain in good condition because there is a thin layer of naturally formed semitransparent mineralised film that is hydrophilic and compact on such rock surface.
In some part under the fine film, knife scratches on the stone from over a thousand years ago are still visible indistinctly. According to solid analysis, natural mineralised film is primarily composed of calcium oxalate monohydrate [Ca(COO)2·H2O] whose process of composition is interfered by microorganism.
There are some advantages of calcium oxalate being a film as follows,
1) In water, especially in acidulous water, the solubility of calcium oxalate is one order of magnitude smaller than normal calcium carbonate.
2) Organically oriented calcium oxalate crystal's particles are fine and compact, which prevent the infiltration of toxic and harmful materials.
3) The system can't be easily broken by external forces in that there is no distinct boundary between calcium carbonate rock and calcium oxalate film.
So, this research project has chosen E. coli that has been transcribed with lipase to clean some stone relics that are covered with oil; then, we use E. coli, with transcription, that can secrete oxalate acid upon the stone relics. Therefore, there forms calcium oxalate to be protection film in order to preserve lithic relics.
If we use chemical methods to generate calcium oxalate, we have to immerse the lithic relics in the chemical liquid; however, it is not suitable for the preservation of large-scale outdoor lithic relics. In fact, the natural existing calcium oxalate comes from the reaction of oxalate acid generated by microorganism and calcium compound on the surface of stone. This research project will utilise genetic engineering methods to "graft" the gene that generates calcium oxalate into Escherichia coli (E. coil).
Then oxalate acid reacts with calcium lithic relics to generate calcium oxalate, which stimulates its natural formation process. Finally, there forms a fine layer to protect the lithic relics.
Procedures in details
4 实验过程
4.1 实验试剂及仪器
4.1.1 目的基因 草酰乙酸乙酰基水解酶(oxaloacetate acetyl hydrolase 简写OAH)是本实验所使用的目的基因片段,主要功效是在表达时将草酰乙酸转化为草酸,草酸进而于钙结合生成草酸钙起到文物保护的作用。此目的基因片段从基因库中查的基因序列后由美国IDT公司直接合成得到。
4.1.2 菌种和质粒 本实验使用的质粒为pET-28a,使用的大肠杆菌为TOP-10感受态细胞,购于天根生化科技有限公司
4.1.3 酶与试剂 实验所使用的限制性内切酶购于NEB,连接酶购于天根生化,DNA纯化试剂盒以及质粒提取试剂盒均购于天根生化。
4.1.4 PCR引物 根据OAH以测得的序列,由Invitrogen合成了如下引物:
上游引物:5’-ATCATCTTGCATATGGCGTCCACCATTGCTGTT-3’(划线部分为Nde i酶切位点)
下游引物:5’-TTTCCTTTTGCGGCCGCTTAG-3’(划线部分为Not i酶切位点)
4.1.5 培养基 实验使用的培养基为LB,以配备200ml的量为例,需要如下材料:胰蛋白胨2g,酵母提取物1g,NaCl 2g,加水至200ml。
实验使用的培养板加入卡那抗性,一配备400ml为例,需要以下材料:胰蛋白胨 4g,酵母提取物 2g,NaCl 4g,Agar琼脂粉6g,卡那霉素 1.2ml
4.2 实验步骤
4.2.1 PCR扩增 因为目的基因片段的G-C碱基对含量很高,所以PCR扩增时需使用GC Buffer,参照Takara产品目录配200ul体系:GC buffer 100ul,dntp mixture 16ul, 前引物 4ul, 后引物 4ul,DNA模板 4ul, Taq 2ul, H2O 70ul。用60℃-70℃进行梯度扩增,寻找引物合适的退火温度。发现在第六条带最亮,再按照上述配成新的体系在适宜温度下进行PCR扩增,得到加上接头的基因片段
4.2.2 做完PCR之后,因有较多的杂条带,故将所有扩增得到的产物跑胶,将所需片段切胶纯化。
4.2.3 进行酶切,分别在目的基因片段和载体pET-28a中加入Not I 和 Nde I 的限制性内切酶。
4.2.4 进行连接并设置对照组。实验组:大片段+小片段+T4 DNA Ligase+Buffer
对照组:大片段 在16℃的环境下连接过夜。
4.2.5 使用DNA纯化试剂盒提取连接后产物,转化进TOP-10感受态细胞,涂在配置好的带卡那抗性的培养板上,放在37℃的恒温环境下培养。
4.2.6 根据菌落的生长情况,将实验组的细菌进行质粒提取,并再次用上述的两种酶进行酶切后跑胶,如果跑胶后大片段和小片段的位置正确,则连接完成。
4.2.7 测序。
4.3 实验结果
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