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A plug‐and‐play pathway refactoring workflow for natural product research in Escherichia coliEscherichia coli and Saccharomyces cerevisiaeSaccharomyces cerevisiae

机译：用于自然产品研究的即插即用途径重构工作流程 escherichia coli escherichia coli和 saccharomyces酿酒酵母酿酒酵母酿酒酵母

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ABSTRACT <section xml:id="bit26309-sec-0001"> > Pathway refactoring serves as an invaluable synthetic biology tool for natural product discovery, characterization, and engineering. However, the complicated and laborious molecular biology techniques largely hinder its application in natural product research, especially in a high‐throughput manner. Here we report a plug‐and‐play pathway refactoring workflow for high‐throughput, flexible pathway construction, and expression in both Escherichia coli and Saccharomyces cerevisiae . Biosynthetic genes were firstly cloned into pre‐assembled helper plasmids with promoters and terminators, resulting in a series of expression cassettes. These expression cassettes were further assembled using Golden Gate reaction to generate fully refactored pathways. The inclusion of spacer plasmids in this system would not only increase the flexibility for refactoring pathways with different number of genes, but also facilitate gene deletion and replacement. As proof of concept, a total of 96 pathways for combinatorial carotenoid biosynthesis were built successfully. This workflow should be generally applicable to different classes of natural products produced by various organisms. Biotechnol. Bioeng. 2017;114: 1847–1854. ? 2017 Wiley Periodicals, Inc. </section> </abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span><Abstract XMLNS =“http://www.wiley.com/namespaces/wiley”type =“main”xml：lang =“en”> <title type =“main”>抽象</ title> <section XML：ID =“Bit26309-SEC-0001”> <P>路径重构用作天然产品发现，表征和工程的无价合成生物工具。然而，复杂和艰苦的分子生物学技术在很大程度上阻碍了其在天然产品研究中的应用，尤其是高吞吐量。在这里，我们报告了一种用于高通量，柔性路径结构的即插即用路径重构工作流程，以及大肠杆菌Coli </ i>和酿酒酵ecesiae</ i>。首先将生物合成基因克隆到具有启动子和终止子的预组装辅助质粒中，得到一系列表达盒。使用金栅反应进一步组装这些表达盒以产生完全重构的途径。在该系统中包含间隔质粒不仅可以增加重构途径的灵活性，而且还促进基因缺失和更换。作为概念证明，共成功建立了组合类胡萝卜素生物合成的96个途径。这种工作流程通常适用于各种生物产生的不同类别的天然产物。 Biotechnol。生物。 2017; 114：1847-1854。还2017 Wiley期刊，Inc。</ p> </ section> </ abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div class="record"> <h2 class="all_title" id="enpatent33" >著录项</h2> <ul> <li> <span class="lefttit">来源</span> <div style="width: 86%;vertical-align: text-top;display: inline-block;"> <a href='/journal-foreign-15017/'>《Biotechnology and Bioengineering》</a> <b style="margin: 0 2px;">|</b><span>2017年第8期</span><b style="margin: 0 2px;">|</b><span>共8页</span> </div> </li> <li> <div class="author"> <span class="lefttit">作者</span> <p id="fAuthorthree" class="threelineshidden zhankaihshouqi"> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ren Hengqian&option=202" target="_blank" rel="nofollow">Ren Hengqian;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hu Pingfan&option=202" target="_blank" rel="nofollow">Hu Pingfan;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zhao Huimin&option=202" target="_blank" rel="nofollow">Zhao Huimin;</a> </p> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zkzz" style="display: none;">展开▼</span> </div> </li> <li> <div style="display: flex;"> <span class="lefttit">作者单位</span> <div style="position: relative;margin-left: 3px;max-width: 639px;"> <div class="threelineshidden zhankaihshouqi" id="fOrgthree"> <p>Department of Chemical and Biomolecular EngineeringUniversity of Illinois at Urbana‐ChampaignUrbana Illinois 61801;</p> <p>Department of Chemical and Biomolecular EngineeringUniversity of Illinois at Urbana‐ChampaignUrbana Illinois 61801;</p> <p>Department of Chemical and Biomolecular EngineeringUniversity of Illinois at Urbana‐ChampaignUrbana Illinois 61801;</p> </div> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zhdw" style="display: none;">展开▼</span> </div> </div> </li> <li > <span class="lefttit">收录信息</span> <span style="width: 86%;vertical-align: text-top;display: inline-block;"></span> </li> <li> <span class="lefttit">原文格式</span> <span>PDF</span> </li> <li> <span class="lefttit">正文语种</span> <span>eng</span> </li> <li> <span class="lefttit">中图分类</span> <span><a href="https://www.zhangqiaokeyan.com/clc/180.html" title="生物工程学（生物技术）">生物工程学（生物技术）;</a></span> </li> <li class="antistop"> <span class="lefttit">关键词</span> <p style="width: 86%;vertical-align: text-top;"> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=natural products&option=203" rel="nofollow">natural products;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=biosynthetic gene clusters&option=203" rel="nofollow">biosynthetic gene clusters;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=pathway refactoring&option=203" rel="nofollow">pathway refactoring;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=synthetic biology&option=203" rel="nofollow">synthetic biology;</a> </p> <div class="translation"> 机译：天然产品;生物合成基因集群;途径重构;合成生物学; </div> </li> </ul> </div> </div> <div class="literature cardcommon"> <div class="similarity "> <h3 class="all_title" id="enpatent66">相似文献</h3> 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