Engineering Escherichia coli for enhanced sensitivity to the autoinducer‐2 quorum sensing signal

Stephens Kristina; Zargar Amin; Emamian Milad; Abutaleb Nadia; Choi Erica; Quan David N.; Payne Gregory; Bentley William E.

摘要

Abstract > The autoinducer‐2 (AI‐2) quorum sensing system is involved in a range of population‐based bacterial behaviors and has been engineered for cell–cell communication in synthetic biology systems. Investigation into the cellular mechanisms of AI‐2 processing has determined that overexpression of uptake genes increases AI‐2 uptake rate, and genomic deletions of degradation genes lowers the AI‐2 level required for activation of reporter genes. Here, we combine these two strategies to engineer an Escherichia coli strain with enhanced ability to detect and respond to AI‐2. In an E. coli strain that does not produce AI‐2, we monitored AI‐2 uptake and reporter protein expression in a strain that overproduced the AI‐2 uptake or phosphorylation units LsrACDB or LsrK, a strain with the deletion of AI‐2 degradation units LsrF and LsrG, and an “enhanced” strain with both overproduction of AI‐2 uptake and deletion of AI‐2 degradation elements. By adding up to 40?μM AI‐2 to growing cell cultures, we determine that this “enhanced” AI‐2 sensitive strain both uptakes AI‐2 more rapidly and responds with increased reporter protein expression than the others. This work expands the toolbox for manipulating AI‐2 quorum sensing processes both in native environments and for synthetic biology applications. </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 Type =“Main”XML：Lang =“en”> <标题类型=“main”>抽象</ title> > AutoInucucer-2（AI-2）仲裁传感系统涉及一系列基于群体的细菌行为，并且已经为合成生物系统中的细胞 - 细胞通信设计。研究AI-2处理的细胞机制已经确定了摄取基因的过表达增加了AI-2摄取率，并且降解基因的基因组缺失降低了对报告基因的激活所需的AI-2水平。在这里，我们将这两种策略与工程师合并大肠杆菌</ i> 应变具有增强的检测和响应AI-2的能力。在A. e。 Coli </ I> 不产生Ai-2的菌株，我们监测AI-2摄取和报告蛋白表达，其菌株过度引起的AI-2摄取或磷酸化单元LSRACDB或LSRK，一种缺失AI-2降解单元LSRF和LSRG的菌株和“增强”菌株具有过量生产的AI-2吸收和缺失Ai-2降解元素。通过增长40μm的Im-2至生长细胞培养物，我们确定该“增强”Ai-2敏感菌株均匀，均匀的AI-2更快，并响应报告蛋白表达的增加而不是其他的。这项工作扩展了工具箱，用于操纵本机环境和合成生物应用中的AI-2仲裁感测过程。 </ p> </摘要> </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-15005/'>《Biotechnology Progress》</a> <b style="margin: 0 2px;">|</b><span>2019年第6期</span><b style="margin: 0 2px;">|</b><span>共6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=Stephens Kristina&option=202" target="_blank" rel="nofollow">Stephens Kristina;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zargar Amin&option=202" target="_blank" rel="nofollow">Zargar Amin;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Emamian Milad&option=202" target="_blank" rel="nofollow">Emamian Milad;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Abutaleb Nadia&option=202" target="_blank" rel="nofollow">Abutaleb Nadia;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Choi Erica&option=202" target="_blank" rel="nofollow">Choi Erica;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Quan David N.&option=202" target="_blank" rel="nofollow">Quan David N.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Payne Gregory&option=202" target="_blank" rel="nofollow">Payne Gregory;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Bentley William E.&option=202" target="_blank" rel="nofollow">Bentley William E.;</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>Fischell Department of BioengineeringUniversity of MarylandCollege Park Maryland;</p> <p>Fischell Department of BioengineeringUniversity of MarylandCollege Park Maryland;</p> <p>Fischell Department of BioengineeringUniversity of MarylandCollege Park Maryland;</p> <p>Fischell Department of BioengineeringUniversity of MarylandCollege Park Maryland;</p> <p>Fischell Department of BioengineeringUniversity of MarylandCollege Park Maryland;</p> <p>Fischell Department of BioengineeringUniversity of MarylandCollege Park Maryland;</p> <p>Institute for Bioscience and Biotechnology Research University of MarylandCollege Park Maryland;</p> <p>Fischell Department of BioengineeringUniversity of MarylandCollege Park Maryland;</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/15.html" title="生物科学">生物科学;</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=AI‐2&option=203" rel="nofollow">AI‐2;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=autoinduction&option=203" rel="nofollow">autoinduction;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=quorum sensing&option=203" rel="nofollow">quorum sensing;</a> </p> <div class="translation"> 机译：AI-2：自动化;他们的感觉; 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Peter Balazovic</a> <span> <a href="/journal-cn-8756/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 今日电子 </a> </span> <span> . 2009</span><span>,第009期</span> </span> </div> </li> <li> <div> <b>6. </b><a class="enjiyixqcontent" href="/academic-conference-cn_meeting-12854_thesis/02022753070.html">基于纳米免疫磁颗粒和金生长信号放大的大肠杆菌O157∶H7QCM传感器检测技术</a> <b>[C]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=李君文&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 李君文</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=谌志强&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,谌志强</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=王景峰&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,王景峰</a> <span> <a href="/conference-cn-12854/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 2010中国人兽共患病学术交流会 </a> <span> <span> . 2010</span> </span> </div> </li> <li> <div> <b>7. </b><a class="enjiyixqcontent" href="/academic-degree-domestic_mphd_thesis/020313516653.html">过表达Beclin1诱导的自噬提高了MG63对Gemcitabine的抵抗与Wnt信号通路关系的研究</a> <b>[A] </b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=陶昊&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 陶昊</a> <span> . 2017</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/061203380466.html">对4-HPPD抑制剂的抵抗性或敏感性提高了的植物</a> <b>[P]</b> . <span> 中国专利： CN105861524B </span> <span> . 2019.07.02</span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/061202027792.html">对4-HPPD抑制剂的抵抗性或敏感性提高了的植物</a> <b>[P]</b> . <span> 中国专利： CN103403165B </span> <span> . 2016.05.04</span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130400497340.html">GENE-THERAPEUTIC DNA-VECTOR BASED ON GENE-THERAPEUTIC DNA-VECTOR VTVAF17, CARRYING TARGET GENE SELECTED FROM GROUP OF GENES ANG, ANGPT1, VEGFA, FGF1, HIF1Α, HGF, SDF1, KLK4, PDGFC, PROK1, PROK2 TO INCREASE EXPRESSION LEVEL OF SAID TARGET GENES, METHOD FOR PRODUCTION AND USE THEREOF, STRAIN ESCHERICHIA COLI SCS110-AF/VTVAF17-ANG, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-ANGPT1, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-VEGFA, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-FGF1, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-HIF1Α, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-HGF, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-SDF1, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-KLK4, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-PDGFC, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-PROK1, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-PROK2, CARRYING GENE-THERAPEUTIC DNA VECTOR, METHOD FOR PRODUCTION THEREOF, METHOD FOR INDUSTRIAL PRODUCTION OF GENE-THERAPEUTIC DNA VECTOR</a> <b>[P]</b> . <span> 外国专利：  RU2730664C2 </span> <span> . 2020-08-24</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：基于基因治疗DNA载体VTVAF17的基因治疗DNA载体，携带选自基因ANG，ANGPT1，VEGFA，FGF1，HIF1α，HGF，SDF1，KLK4，PDGFC，PROK1，PROK2表达的靶基因所述的靶标基因，其生产和使用方法，应变大肠埃希氏菌SCS110-AF / VTVAF17-ANG或大肠埃希氏菌SCS110-AF / VTVAF17-ANGPT1或大肠埃希氏菌SCS110-AF / VTVAF17-VEGFA或大肠埃希氏菌SCS110-AF / VTVAF17-FGF1，或大肠埃希氏菌SCS110-AF / VTVAF17-HIF1A，或大肠埃希氏菌COLI SCS110-AF / VTVAF17-HGF，或大肠埃希氏菌SCS110-AF / VTVAF17-SDF1，或大肠埃希氏菌SCS110-AF / VTVAF17-KLK4，大肠埃希氏菌SCS110-AF / VTVAF17-PDGFC或大肠埃希氏菌SCS110-AF / VTVAF17-PROK2，携带基因-治疗性DNA载体，生产方法，工业生产方法-治疗性DNA载体 </span> </p> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/patent-detail/06130400493335.html">GENE-THERAPEUTIC DNA VECTOR BASED ON THE GENE-THERAPEUTIC DNA VECTOR GDTT1_8NAS12, CARRYING THE TARGET GENE SELECTED FROM A GROUP OF GENES DDC, IL10, IL13, IFNB1, TNFRSF4, TNFSF10, BCL2, HGF, IL2 TO INCREASE THE EXPRESSION LEVEL OF SAID TARGET GENES, A METHOD FOR PRODUCTION AND USE THEREOF, A STRAIN ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-DDC OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-IL10 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-IL13 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-IFNB1 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-TNFRSF4 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-TNFSF10 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-BCL2 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-HGF OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-IL2, CARRYING A GENE-THERAPEUTIC DNA VECTOR, METHOD FOR PRODUCTION THEREOF, A METHOD FOR INDUSTRIAL PRODUCTION OF A GENE-THERAPEUTIC DNA VECTOR</a> <b>[P]</b> . <span> 外国专利：  RU2734726C1 </span> <span> . 2020-10-22</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：基于基因治疗DNA矢量GDTT1_8NAS12的基因治疗DNA矢量，携带从一组基因中选择的目标基因DDC，IL10，IL13，IFNB1，TNFRSF4，TNFSF10，BCL2，HGF，IL2可以增加表达水平基因，一种生产和使用其的方法，应变大肠埃希氏菌JM110-NAS / GDTT1_8NAS12-DDC或大肠埃希氏菌JM110-NAS / GDTT1_8NAS12-IL10或大肠埃希氏菌JM110-NAS / GDTT1_8NAS12-IL13或大肠埃希氏菌COLI JM110-NAS / GDTT1_8NAS12-IL13 IFNB1或ESCHERICHIA COLI JM110-NAS / GDTT1_8NAS12-TNFRSF4或ESCHERICHIA COLI JM110-NAS / GDTT1_8NAS12-TNFSF10或ESCHERICHIA COLI JM110-NAS / GDTT1_8NAS12-BCL2或ESCHERICHIA COLI JM110-NAS / GDTT1_8NAS12 IL2，携带基因治疗性DNA载体，其生产方法，工业生产基因治疗性DNA载体的方法 </span> </p> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/patent-detail/06130400537005.html">Gene therapeutic DNA vector based on VTvaf17 gene therapeutic DNA vector carrying a target gene selected from the group of genes ANG, ANGPT1, VEGFA, FGF1, HIF1α, HGF, SDF1, KLK4, PDGFC, PROK1, PROK2 to increase the expression level of these target genes, method its preparation and use, Escherichia coli strain SCS110-AF / VTvaf17-ANG or Escherichia coli SCS110-AF / VTvaf17-ANGPT1 or Escherichia coli SCS110-AF / VTvaf17-VEGFA or Escherichia coli SCS110-AF / VTvaf17-Fichia-SC110 AF / VTvaf17-HIF1α or Escherichia coli SCS110-AF / VTvaf17-HGF or Escherichia coli SCS110-AF / VTvaf17-SDF1 or Escherichia coli SCS110-AF / VTvaf17-KLK4 or Escherichia coli SCS110-AF / VTviFi10 SCF110 AF / VTvaf17-PROK1 or Escherichia coli SCS110-AF / VTvaf17-PROK2 carrying a gene therapy DNA vector, a method for its preparation, a method for the industrial production of a gene therapy DNA vector</a> <b>[P]</b> . <span> 外国专利：  RU2018147085A </span> <span> . 2020-06-29</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：基于VTvaf17基因治疗DNA载体的基因治疗DNA载体，其携带选自ANG，ANGPT1，VEGFA，FGF1，HIF1α，HGF，SDF1，KLK4，PDGFC，PROK1，PROK2的靶基因以增加这些靶标的表达水平基因，方法的制备和使用，大肠杆菌菌株SCS110-AF / VTvaf17-ANG或大肠杆菌SCS110-AF / VTvaf17-ANGPT1或大肠杆菌SCS110-AF / VTvaf17-VEGFA或大肠杆菌SCS110-AF / VTvaf17-Fichia-SC110 AF /VTvaf17-HIF1α或大肠杆菌SCS110-AF / VTvaf17-HGF或大肠杆菌SCS110-AF / VTvaf17-SDF1或大肠杆菌SCS110-AF / VTvaf17-KLK4或大肠杆菌SCS110-AF / VTviFi10 S1携带基因治疗DNA载体的大肠杆菌SCS110-AF / VTvaf17-PROK2，其制备方法，工业生产基因治疗DNA载体的方法 </span> </p> </li> </ul> </div> </div> </div> <div class="theme cardcommon" style="overflow: auto;display:none"> <h3 class="all_title" id="enpatent55">相关主题</h3> <ul id="subject"> </ul> </div> </div> </div> </div> <div class="right rightcon"> <div class="details_img cardcommon clearfix" style="margin-bottom: 10px;display:none;" > </div> </div> </div> <div id="thesis_get_original1" class="downloadBth" style="bottom: 19px;z-index: 999;" onclick="ywcd('0704021880228','4',7,2,1,'',this,24)" class="delivery" prompt="010401" title="通过人工服务将文献原文发送至邮箱" >获取原文</div> <div class="journalsub-pop-up" style="display: none"> <div class="journal-sub"> <h2>期刊订阅</h2> <img src="https://cdn.zhangqiaokeyan.com/img/loginclose.png" alt="关闭" onclick="$('.journalsub-pop-up').hide()"> <p class="pardon">抱歉，该期刊暂不可订阅，敬请期待！</p> <p class="current">目前支持订阅全部北京大学中文核心（2020）期刊目录。</p> <div style="display: flex;margin-top: 69px;justify-content: 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