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<h1 id="分子生物学期末考试-回忆版（25-26秋）"><a href="#分子生物学期末考试-回忆版（25-26秋）" class="headerlink" title="分子生物学期末考试 回忆版（25-26秋）"></a>分子生物学期末考试 回忆版（25-26秋）</h1><h2 id="名词解释"><a href="#名词解释" class="headerlink" title="名词解释"></a>名词解释</h2><p>操纵子、断裂基因、基因组、顺式作用元件、反式作用因子、RNA干扰、复制叉、复性、表观遗传</p>
<h2 id="问答题"><a href="#问答题" class="headerlink" title="问答题"></a>问答题</h2><p>为什么DNA双链比RNA单链更适合作为遗传物质？</p>
<p>哺乳动物如何解决DNA复制末端的稳定性问题？</p>
<p>原核生物强启动子的构成要素</p>
<p>蛋白质生物合成过程中，RNA发挥什么作用？</p>
<p>原核生物、真核生物mRNA转录的不同</p>
<h2 id="论述题"><a href="#论述题" class="headerlink" title="论述题"></a>论述题</h2><p>给出乳糖操纵子图。上面各个字母是什么意思？结构基因三个分别表达什么酶？为什么同时有乳糖和葡萄糖，优先利用葡萄糖？</p>
<p>DNA半保留复制是什么？设计实验证明。至少列举2个利用DNA半保留复制的分子生物学技术，说明原理。</p>
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<h1 id="分子生物学总结"><a href="#分子生物学总结" class="headerlink" title="分子生物学总结"></a>分子生物学总结</h1><h2 id="一、-DNA复制-DNA-Replication"><a href="#一、-DNA复制-DNA-Replication" class="headerlink" title="一、 DNA复制 (DNA Replication)"></a>一、 DNA复制 (DNA Replication)</h2><p><strong>核心目标：</strong> 以DNA为模板合成DNA<br><strong>方向：</strong> $5’ \to 3’$</p>
<div class="table-container">
<table>
<thead>
<tr>
<th style="text-align:left">比较项目</th>
<th style="text-align:left"><strong>原核生物 (Prokaryotes)</strong></th>
<th style="text-align:left"><strong>真核生物 (Eukaryotes)</strong></th>
</tr>
</thead>
<tbody>
<tr>
<td style="text-align:left"><strong>发生场所</strong></td>
<td style="text-align:left">细胞质（拟核）</td>
<td style="text-align:left">细胞核、线粒体、叶绿体</td>
</tr>
<tr>
<td style="text-align:left"><strong>发生时间</strong></td>
<td style="text-align:left">细胞分裂期间，整个细胞周期</td>
<td style="text-align:left">细胞周期的 <strong>S期</strong></td>
</tr>
<tr>
<td style="text-align:left"><strong>复制起点</strong></td>
<td style="text-align:left"><strong>单一起点 (oriC)</strong></td>
<td style="text-align:left"><strong>多起点</strong> (复制子)，为了在短时间内复制巨大基因组</td>
</tr>
<tr>
<td style="text-align:left"><strong>关键酶</strong></td>
<td style="text-align:left"><strong>解旋酶 (DnaB)</strong>：解开双链<br><strong>引发酶 (DnaG)</strong>：合成RNA引物<br><strong>DNA Pol III</strong>：<strong>主力军</strong>，负责大部分延伸<br><strong>DNA Pol I</strong>：切除引物并填补空缺<br><strong>DNA连接酶</strong>：连接冈崎片段</td>
<td style="text-align:left"><strong>解旋酶 (MCM复合物)</strong><br><strong>DNA Pol $\alpha$</strong>：含引发酶活性，合成引物<br><strong>DNA Pol $\delta$</strong>：主要负责<strong>后随链</strong>合成<br><strong>DNA Pol $\varepsilon$</strong>：主要负责<strong>前导链</strong>合成<br><strong>FEN1/RNase H</strong>：切除引物</td>
</tr>
<tr>
<td style="text-align:left"><strong>过程：起始</strong></td>
<td style="text-align:left">DnaA蛋白识别oriC，解旋酶解开双链，SSB结合单链。</td>
<td style="text-align:left">ORC识别起点，<strong>细胞周期调控</strong>因子(Cdc6等)组装前复制复合物。</td>
</tr>
<tr>
<td style="text-align:left"><strong>过程：延伸</strong></td>
<td style="text-align:left">同样是半不连续复制（前导链/后随链）。</td>
<td style="text-align:left">同样是半不连续复制。冈崎片段比原核短。伴随<strong>核小体</strong>的解聚与重组。</td>
</tr>
<tr>
<td style="text-align:left"><strong>过程：终止</strong></td>
<td style="text-align:left"><strong>Ter序列</strong>结合Tus蛋白阻滞复制叉；<strong>Topo IV</strong> 解开连环体。</td>
<td style="text-align:left">复制叉相遇融合。<strong>端粒酶</strong>解决线性染色体末端缩短问题（爬行模型）。</td>
</tr>
</tbody>
</table>
</div>
<hr>
<h2 id="二、-转录-Transcription"><a href="#二、-转录-Transcription" class="headerlink" title="二、 转录 (Transcription)"></a>二、 转录 (Transcription)</h2><p><strong>核心目标：</strong> 以DNA为模板合成RNA<br><strong>方向：</strong> $5’ \to 3’$</p>
<div class="table-container">
<table>
<thead>
<tr>
<th style="text-align:left">比较项目</th>
<th style="text-align:left"><strong>原核生物 (Prokaryotes)</strong></th>
<th style="text-align:left"><strong>真核生物 (Eukaryotes)</strong></th>
</tr>
</thead>
<tbody>
<tr>
<td style="text-align:left"><strong>发生场所</strong></td>
<td style="text-align:left">细胞质</td>
<td style="text-align:left">细胞核（转录与翻译<strong>时空分隔</strong>）</td>
</tr>
<tr>
<td style="text-align:left"><strong>酶</strong></td>
<td style="text-align:left"><strong>只有一种 RNA聚合酶</strong><br>全酶 = 核心酶 ($\alpha_2\beta\beta’\omega$) + <strong>$\sigma$因子</strong></td>
<td style="text-align:left"><strong>三种 RNA聚合酶</strong><br><strong>Pol I</strong> $\to$ rRNA<br><strong>Pol II</strong> $\to$ <strong>mRNA</strong> (核心)<br><strong>Pol III</strong> $\to$ tRNA, 5S rRNA</td>
</tr>
<tr>
<td style="text-align:left"><strong>识别机制</strong></td>
<td style="text-align:left"><strong>$\sigma$ 因子</strong> 直接识别启动子 (-10区, -35区)。</td>
<td style="text-align:left">RNA聚合酶不能直接识别，需<strong>通用转录因子 (GTFs)</strong> (如TFIID中的TBP) 识别启动子 (<strong>TATA box</strong>)。</td>
</tr>
<tr>
<td style="text-align:left"><strong>过程：起始</strong></td>
<td style="text-align:left">全酶结合启动子 $\to$ 形成闭合复合物 $\to$ 开放复合物 $\to$ 合成第一个磷酸二酯键 $\to$ $\sigma$因子脱落。</td>
<td style="text-align:left">TBP结合TATA盒 $\to$ 招募TFIIB/F/E/H $\to$ 形成PIC $\to$ <strong>TFIIH磷酸化Pol II的CTD</strong> $\to$ 启动延伸。</td>
</tr>
<tr>
<td style="text-align:left"><strong>过程：延伸</strong></td>
<td style="text-align:left">核心酶沿模板移动，形成转录泡。<strong>转录翻译偶联</strong>。</td>
<td style="text-align:left">Pol II 延伸，伴随5’端加帽。<strong>无转录翻译偶联</strong>。</td>
</tr>
<tr>
<td style="text-align:left"><strong>过程：终止</strong></td>
<td style="text-align:left">1. <strong>不依赖$\rho$因子</strong>：形成茎环结构+Poly U。<br>2. <strong>依赖$\rho$因子</strong>：$\rho$蛋白追赶酶，解开杂合链。</td>
<td style="text-align:left">机制复杂，通常与 <strong>3’端加工（加尾）</strong> 偶联。AAUAAA信号介导切除和加尾。</td>
</tr>
<tr>
<td style="text-align:left"><strong>转录后加工</strong></td>
<td style="text-align:left">极少（mRNA直接翻译）。</td>
<td style="text-align:left"><strong>复杂</strong>：<br>1. <strong>5’加帽</strong> (m7G)<br>2. <strong>3’加尾</strong> (Poly A)<br>3. <strong>剪接</strong> (去除内含子)</td>
</tr>
<tr>
<td style="text-align:left"><strong>产物形式</strong></td>
<td style="text-align:left"><strong>多顺反子</strong> (一条mRNA编码多个蛋白)</td>
<td style="text-align:left"><strong>单顺反子</strong> (一条mRNA编码一个蛋白)</td>
</tr>
</tbody>
</table>
</div>
<hr>
<h2 id="三、-翻译-Translation"><a href="#三、-翻译-Translation" class="headerlink" title="三、 翻译 (Translation)"></a>三、 翻译 (Translation)</h2><p><strong>核心目标：</strong> 以mRNA为模板合成蛋白质<br><strong>方向：</strong> 肽链 N端 $\to$ C端</p>
<div class="table-container">
<table>
<thead>
<tr>
<th style="text-align:left">比较项目</th>
<th style="text-align:left"><strong>原核生物 (Prokaryotes)</strong></th>
<th style="text-align:left"><strong>真核生物 (Eukaryotes)</strong></th>
</tr>
</thead>
<tbody>
<tr>
<td style="text-align:left"><strong>核糖体</strong></td>
<td style="text-align:left"><strong>70S</strong> (30S小亚基 + 50S大亚基)</td>
<td style="text-align:left"><strong>80S</strong> (40S小亚基 + 60S大亚基)</td>
</tr>
<tr>
<td style="text-align:left"><strong>起始氨基酸</strong></td>
<td style="text-align:left"><strong>fMet</strong> (N-甲酰甲硫氨酸)</td>
<td style="text-align:left"><strong>Met</strong> (甲硫氨酸)</td>
</tr>
<tr>
<td style="text-align:left"><strong>起始tRNA</strong></td>
<td style="text-align:left">$tRNA_f^{Met}$</td>
<td style="text-align:left">$tRNA_i^{Met}$</td>
</tr>
<tr>
<td style="text-align:left"><strong>识别信号</strong></td>
<td style="text-align:left"><strong>SD序列</strong> (AGGAGG)，与16S rRNA互补配对。</td>
<td style="text-align:left"><strong>5’ 帽子结构</strong> (m7G) 和 <strong>Kozak序列</strong>。</td>
</tr>
<tr>
<td style="text-align:left"><strong>过程：起始</strong></td>
<td style="text-align:left">IF1/2/3因子协助。30S亚基直接结合SD序列 $\to$ fMet-tRNA进入P位 $\to$ 50S结合。</td>
<td style="text-align:left"><strong>扫描模型</strong>：eIF4F识别帽子 $\to$ 40S亚基结合 $\to$ 扫描寻找第一个AUG $\to$ 60S结合。</td>
</tr>
<tr>
<td style="text-align:left"><strong>过程：延伸</strong></td>
<td style="text-align:left">1. <strong>进位</strong>：EF-Tu (GTP)<br>2. <strong>成肽</strong>：23S rRNA (核酶)<br>3. <strong>移位</strong>：EF-G (GTP)</td>
<td style="text-align:left">1. <strong>进位</strong>：eEF1$\alpha$ (GTP)<br>2. <strong>成肽</strong>：28S rRNA (核酶)<br>3. <strong>移位</strong>：eEF2 (GTP)</td>
</tr>
<tr>
<td style="text-align:left"><strong>过程：终止</strong></td>
<td style="text-align:left"><strong>RF1</strong> (认UAA/UAG) 或 <strong>RF2</strong> (认UAA/UGA)。</td>
<td style="text-align:left">只有 <strong>eRF1</strong> (识别所有3种终止密码子)。</td>
</tr>
<tr>
<td style="text-align:left"><strong>多聚核糖体</strong></td>
<td style="text-align:left">有</td>
<td style="text-align:left">有</td>
</tr>
</tbody>
</table>
</div>
<hr>
<h2 id="四、-极简速记口诀"><a href="#四、-极简速记口诀" class="headerlink" title="四、 极简速记口诀"></a>四、 极简速记口诀</h2><ol>
<li><p><strong>复制</strong>：</p>
<ul>
<li><strong>原核</strong>：单起点，Pol III 干活，Pol I 修补，冈崎片段长，引物切除靠 Pol I。</li>
<li><strong>真核</strong>：多起点，Pol $\delta/\varepsilon$ 干活，$\alpha$ 起始，有端粒酶，有核小体，冈崎片段短。</li>
</ul>
</li>
<li><p><strong>转录</strong>：</p>
<ul>
<li><strong>原核</strong>：1种酶，靠 $\sigma$ 认路，边录边译，多顺反子。</li>
<li><strong>真核</strong>：3种酶 (II造mRNA)，靠 TBP 认路，先加帽加尾剪接再出核，单顺反子。</li>
</ul>
</li>
<li><p><strong>翻译</strong>：</p>
<ul>
<li><strong>原核</strong>：70S，SD序列找AUG，首氨基酸带帽子(fMet)。</li>
<li><strong>真核</strong>：80S，帽子扫描找AUG，首氨基酸不带帽(Met)。</li>
</ul>
</li>
</ol>
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