designing

asserts/css/style.css

@@ -420,7 +420,7 @@ body.dark-theme .translate-icon:hover path {
 /* 链接 hover下划线 */
 a {
     position: relative;
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+    font-size: 2rem;
     text-decoration: none;
     font-weight: 600;
     color: var(--link-color);
@@ -618,7 +618,14 @@ p {
     font-weight: 350;
     line-height:150%;
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+.math-equation {
+    display: block;
+    margin: 1.5rem 0;
+    text-align: center;
+    padding: 0.5rem;
+    background-color: #f7f7f7;
+    font-size: 1.2em;
+}
 strong {
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pages/design.html

@@ -232,7 +232,11 @@ <h1 class="leader img-fluid">Design</h1>
             </div>
             <!-- Header Section Content -->
         </div>
-
+        <div class="row">
+            <div class="col-md-3"></div>
+            <img src="../asserts/img/design/design1.png" alt="Figure 1" class="img-fluid col-md-6">
+            <p class="center-aligned-text">Fig 1</p>
+        </div>
         <div class="container">
             <br><br>
             <h2 id="01" class="module">Module One</h2>
@@ -342,6 +346,11 @@ <h3>Antigen Testing</h3>
             <p class="cn d-none">在抗体检测的基础上，对我的系统稍加修改我们就能实现对于指定种类抗原的检测。
                 纳米抗体是在骆驼科动物血液中发现的一种缺失轻链的特殊抗体，即重链抗体。和普通抗体相比，它只包含一个重链可变区（VHH）和两个常规的CH2与CH3区。纳米抗体具有体积小，效能稳定，抗原特异性好，且易于基因改造等优点。除此之外，一个非常吸引人的点是纳米抗体可以在原核表达体系内表达[12]。
             </p>
+            <div class="row">
+                <div class="col-md-3"></div>
+                <img src="../asserts/img/design/design_6.png" alt="Figure 6" class="img-fluid col-md-6">
+                <p class="center-aligned-text">Fig 6</p>
+            </div>
             <p class="en">By fusing the nanobody specific to a particular antigen with Snoopcatcher in a prokaryotic
                 system and connecting it to e-pili containing Snooptag, we can obtain a batch of e-pili capable of
                 detecting the specified antigen.</p>
@@ -367,6 +376,11 @@ <h3 class=>Nucleic Acid Testing</h3>
                 the Halo Tag ligand into the system.</p>
             <p class="cn d-none">若将Snoopcatcher与Halo
                 Tag融合表达,使其与相应配体修饰的单链随机寡核苷酸反应，则最终有望实现寡核苷酸和导电菌毛的相连，并利用随机寡核苷酸与猎物核酸分子的分子杂交而引发的电导率变化检测该核酸分子。</p>
+                <div class="row">
+                    <div class="col-md-3"></div>
+                    <img src="../asserts/img/design/design_7.png" alt="Figure 7" class="img-fluid col-md-6">
+                    <p class="center-aligned-text">Fig 7</p>
+                </div>
             <h2 id="04" class="module">Chassis Microbes</h2>
             <p class="en">The selection of chassis microbes is also a crucial issue in our project. G.metallireducdens.
                 is a strictly anaerobic that is challenging to industrially cultivate with existing technologies. In
@@ -451,6 +465,16 @@ <h2 id="05" class="module">CRISPRi System</h2>
             <p class="cn d-none">
                 将报告质粒与pdCas9-sg共同转入需钠弧菌当中，即可通过检测绿色荧光的强度来判断该sgRNA对于V.PilA的抑制效果。这一报告系统避免了测试CRISPRi工作效果时需反复收获菌毛的问题，同时也避免了菌毛收获过程中操作带来的误差。
             </p>
+            <div class="row">
+                <div class="col-md-1"></div>
+                <img src="../asserts/img/design/design_8_1.png" alt="Figure 8_1" class="img-fluid col-md-4">
+                <div class="col-md-1"></div>
+                <img src="../asserts/img/design/design_8_2.png" alt="Figure 8_2" class="img-fluid col-md-5">
+                <div class="col-md-1"></div>
+                <div class="col-md-1"></div>
+                <img src="../asserts/img/design/design8_3.png" alt="Figure 8_3" class="img-fluid col-md-10">
+                <p class="center-aligned-text">Fig 8</p>
+            </div>
             <h2 id="06" class="module">The Work Flow</h2>
             <p class="en">Our project plan is divided into the following five parts: establishment of CRISPRi testing
                 system and pilus expression system, optimization of CRISPRi, expression and harvest of Geobacter
@@ -496,13 +520,23 @@ <h3 class="cn d-none">菌毛表达质粒 pPilin </h3>
                 metallireducens type IV pilus.</p>
             <p class="cn d-none">pPilin是表达导电菌毛的主要质粒。该质粒以pUC19作为骨架，并以pBAD（BBa_I13453）控制金属还原地杆菌四型菌毛单体PilA（简称为G.PilA）的表达。
             </p>
+            <div class="row">
+                <div class="col-md-3"></div>
+                <img src="../asserts/img/design/design9.png" alt="Figure 9" class="img-fluid col-md-6">
+                <p class="center-aligned-text">Fig 9</p>
+            </div>
             <h3 class="en">CRISPRi plasmid—pdCas9-sg</h3>
             <h3 class="cn d-none">CRISPRi质粒pdCas9-sg</h3>
             <p class="en">pdCas9-sg uses pRSFDuet as a backbone and expresses both spdCas9 and sgRNA. spdCas9
                 transcription is controlled by Ptrc and induced by IPTG, and sgRNA is expressed under the control of the
                 constitutive promoter BBa_J23100.</p>
             <p class="cn d-none">
                 pdCas9-sg以pRSFDuet作为骨架，同时表达spdCas9和sgRNA，其中spdCas9由Ptrc控制转录在IPTG的诱导下表达，sgRNA由组成性启动子BBa_J23100控制表达。</p>
+                <div class="row">
+                    <div class="col-md-3"></div>
+                    <img src="../asserts/img/design/design10.png" alt="Figure 10" class="img-fluid col-md-6">
+                    <p class="center-aligned-text">Fig 10</p>
+                </div>
             <p class="en">We initially used pdCas9-sg1 expressing sgRNA1 ordered from AZENTA. Then to test the
                 inhibitory effect of different sgRNAs, we plan to construct CRISPRi plasmids expressing three other
                 sgRNAs using Gibson assembly. The sgRNA fragments for substitution were ordered from AZENTA, and then
@@ -511,6 +545,11 @@ <h3 class="cn d-none">CRISPRi质粒pdCas9-sg</h3>
             <p class="cn d-none">我们最初使用的为表达sgRNA1的pdCas9-sg1，该质粒由AZENTA公司订购。然后为了测试不同sgRNA的抑制效果，我们计划利用Gibson
                 assembly构建表达另外三个sgRNA的CRISPRi质粒。替换用的sgRNA片段从AZENTA公司订购，然后通过反向PCR将pdCas9-sg进行线性化并添加同源臂，利用gibson
                 assembly进行组装获得pdCas9-sg2, pdCas9-sg3和pdCas9-sg4。</p>
+                <div class="row">
+                    <div class="col-md-3"></div>
+                    <img src="../asserts/img/design/design_11.png" alt="Figure 11" class="img-fluid col-md-6">
+                    <p class="center-aligned-text">Fig 11</p>
+                </div>
             <h3 class="en">reporter plasmid—pGFPmod2</h3>
             <h3 class="cn d-none">报告质粒pGFPmod2</h3>
             <p class="en">The reporter plasmid based on sfGFP uses pUC19 as the backbone. The expression of recombinant
@@ -525,6 +564,12 @@ <h3 class="cn d-none">报告质粒pGFPmod2</h3>
             <p class="cn d-none">
                 基于sfGFP的报告质粒以pUC19作为骨架，由需钠弧菌菌毛单体PilA（V.PilA）的启动子控制重组sfGFP的表达，同时在重组sfGFP的N端连接了一段去除了起始密码子的V.PilA
                 N端序列。我们选择的四个sgRNA均靶向到这V.PilA转录起始位点（TSS）附近，即包括V.PilA的启动子和N端编码序列，因此这一报告质粒可以通过绿色荧光的强度来提示CRISPRi系统的工作效果。我们在设计报告质粒时经过了一系列调整，最终获得了具有较好效果的pGFPmod2版本。</p>
+
+                <div class="row">
+                    <div class="col-md-3"></div>
+                    <img src="../asserts/img/design/design12.png" alt="Figure 12" class="img-fluid col-md-6">
+                    <p class="center-aligned-text">Fig 12</p>
+                </div>
             <h2 id="08" class="module">Purification</h2>
             <p class="en">In order to test the conductivity of pilus and its ability to detect concentrations in the
                 laboratory, we need to harvest large quantities of conductivepilus and make them into conductive sheets.
@@ -538,6 +583,11 @@ <h3 class="en">Shear and filter</h3>
             <h3 class="cn d-none">剪切过滤</h3>
             <p class="en">This method acquires pilus by a purely physical means. We intend to refer to the harvesting method of conductive pilus of Geobacter sulfurreducens to harvest large quantities of conductive pilus assembled from G. pilA expressed by Vibrio natriegens. The pili were first cut from the bacterial surface by mechanical agitation in a waring blender, and the cellular debris was removed before the mixed solution was filtered under nitrogen aeration. Since the molecular weight of this conductive pilus is more than 100kDa, the use of 100kD cut-off PVDF membrane can separate the pili from other proteins with smaller molecular weights. Thus the final solution of pili is obtained. This method can treat a large number of bacteria in a single run, although the purity of the pili obtained is not high.</p>
             <p class="cn d-none">该方法完全通过物理的方式对菌毛进行获取。我们打算参考硫还原地杆菌导电菌毛的收获方法[22]，对需钠弧菌表达的G.PilA组装成为的导电菌毛进行大批量收获。首先通过在waring blender中机械搅拌的方法将菌毛从细菌表面切割下来，去除细胞碎片后再在氮气通气和搅拌下对混合溶液进行过滤。由于该导电菌毛的分子量大于100kDa，利用100kDa截留的PVDF膜可以将菌毛和其他分子量较小的蛋白分开，最终得到菌毛溶液。该方法虽然获得的菌毛纯度不高，但单次可以处理大量的细菌。</p>
+            <div class="row">
+                <div class="col-md-3"></div>
+                <img src="../asserts/img/design/design13.png" alt="Figure 13" class="img-fluid col-md-6">
+                <p class="center-aligned-text">Fig 13</p>
+            </div>
             <h3 class="en">Ni-NTA purification</h3>
             <h3 class="cn d-none">Ni-NTA树脂纯化</h3>
             <p class="en">In addition to the efficient large-scale harvesting method, we have also opted for a