Initiation of Protein Synthesis with Non‐Canonical Amino Acids In?Vivo

Tharp Jeffery M.; Ad Omer; Amikura Kazuaki; Ward Fred R.; Garcia Emma M.; Cate Jamie H. D.; Schepartz Alanna; S?ll Dieter

摘要

Abstract > By transplanting identity elements into E.?coli tRNA fMet , we have engineered an orthogonal initiator tRNA (itRNA Ty2 ) that is a substrate for Methanocaldococcus jannaschii TyrRS. We demonstrate that itRNA Ty2 can initiate translation in?vivo with aromatic non‐canonical amino acids (ncAAs) bearing diverse sidechains. Although the initial system suffered from low yields, deleting redundant copies of tRNA fMet from the genome afforded an E.?coli strain in which the efficiency of non‐canonical initiation equals elongation. With this improved system we produced a protein containing two distinct ncAAs at the first and second positions, an initial step towards producing completely unnatural polypeptides in?vivo. This work provides a valuable tool to synthetic biology and demonstrates remarkable versatility of the E.?coli translational machinery for initiation with ncAAs in vivo. </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> > 通过将同一性元素移植到E.?COLI TRNA中 fmet </ sup> ，我们设计了正交引发剂TRNA（ITRNA ty2 </ sup> ）这是甲烷甘油干癌基因菌属Tyrrs的底物。我们证明了ITRNA ty2 </ sup> 可以在αvivo与芳香族非规范氨基酸（NCAAs）轴承不同的侧链拟合翻译。虽然初始系统患有低产量，但删除了TRNA的冗余副本 fmet </ sup> 来自基因组，得到了一种E.?COLI菌株，其中非规范发起的效率等于伸长率。利用这种改进的系统，我们在第一和第二位置产生了含有两个不同NCAA的蛋白质，旨在在β体内产生完全不自然多肽的初始步骤。这项工作为合成生物学提供了有价值的工具，并展示了E.OSCOLI翻译机械的显着多功能性，用于在体内启动NCAAs。 </ 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-19011/'>《Angewandte Chemie》</a> <b style="margin: 0 2px;">|</b><span>2020年第8期</span><b style="margin: 0 2px;">|</b><span>共5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=Tharp Jeffery M.&option=202" target="_blank" rel="nofollow">Tharp Jeffery M.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ad Omer&option=202" target="_blank" rel="nofollow">Ad Omer;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Amikura Kazuaki&option=202" target="_blank" rel="nofollow">Amikura Kazuaki;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ward Fred R.&option=202" target="_blank" rel="nofollow">Ward Fred R.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Garcia Emma M.&option=202" target="_blank" rel="nofollow">Garcia Emma M.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Cate Jamie H. D.&option=202" target="_blank" rel="nofollow">Cate Jamie H. D.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Schepartz Alanna&option=202" target="_blank" rel="nofollow">Schepartz Alanna;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=S?ll Dieter&option=202" target="_blank" rel="nofollow">S?ll Dieter;</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 Molecular Biophysics and BiochemistryYale UniversityNew Haven CT 06520 USA;</p> <p>Department of ChemistryYale UniversityNew Haven CT 06520 USA;</p> <p>Department of Molecular Biophysics and BiochemistryYale UniversityNew Haven CT 06520 USA;</p> <p>Department of Molecular and Cell BiologyUniversity of CaliforniaBerkeley CA 94720 USA;</p> <p>Department of Molecular Biophysics and BiochemistryYale UniversityNew Haven CT 06520 USA;</p> <p>Department of Molecular and Cell BiologyUniversity of CaliforniaBerkeley CA 94720 USA;</p> <p>Department of ChemistryYale UniversityNew Haven CT 06520 USA;</p> <p>Department of Molecular Biophysics and BiochemistryYale UniversityNew Haven CT 06520 USA;</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/1188.html" title="应用化学">应用化学;</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=chemical biology&option=203" rel="nofollow">chemical biology;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=genetic code expansion&option=203" rel="nofollow">genetic code expansion;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=non-canonical amino acids&option=203" rel="nofollow">non-canonical amino acids;</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> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=translation initiation&option=203" rel="nofollow">translation initiation;</a> </p> <div class="translation"> 机译：化学生物学;遗传规范扩张;非典型氨基酸;合成生物学;翻译启动; </div> </li> </ul> </div> </div> <div class="literature cardcommon"> <div class="similarity "> <h3 class="all_title" id="enpatent66">相似文献</h3> <div 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href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=叶元土&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,叶元土</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=林仕梅&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,林仕梅</a> <span> <a href="/journal-cn-7204/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 水产学报 </a> </span> <span> . 2002</span><span>,第001期</span> </span> </div> </li> <li> <div> <b>6. </b><a class="enjiyixqcontent" href="/academic-conference-cn_meeting-23236_thesis/02022827868.html">谷氨酰胺联合生长激素对脓毒症大鼠血浆氨基酸谱和骨胳肌蛋白质合成代谢的影响</a> <b>[C]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=刘敏丰&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 刘敏丰</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=高志光&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,高志光</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=秦环龙&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,秦环龙</a> <span> <a href="/conference-cn-23236/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 第十届全国临床营养支持学术会议 </a> <span> <span> . 2006</span> </span> </div> </li> <li> <div> <b>7. </b><a class="enjiyixqcontent" href="/academic-degree-domestic_mphd_thesis/020313491685.html">阴外动脉灌注氨基酸和脂肪酸对奶牛乳腺蛋白质合成和氨基酸摄取规律的影响</a> <b>[A] </b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=王新朋&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 王新朋</a> <span> . 2015</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/06120107861461.html">含有非天然氨基酸和多肽的组合物、涉及非天然氨基酸和多肽的方法以及非天然氨基酸和多肽的用途</a> <b>[P]</b> . <span> 中国专利： CN105384807A </span> <span> . 2016-03-09</span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/061201817646.html">含有非天然氨基酸和多肽的组合物、涉及非天然氨基酸和多肽的方法以及非天然氨基酸和多肽的用途</a> <b>[P]</b> . <span> 中国专利： CN101448512B </span> <span> . 2015.11.25</span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130416609308.html">RECOMBINANT PLASMID DNA pPA-OPRF-ETA, WHICH CODES SYNTHESIS OF HYBRID RECOMBINANT PROTEIN, WHICH INCLUDES AMINO-ACID SEQUENCES OF PROTEIN F OF OUTER MEMBRANE AND EXOTOXIN A PSEUDOMONAS aeruginosa, DEFORMATION OF ESCHERICHIA coli PA-OPRF-ETA - PRODUCER OF HYBRID RECOMBINANT PROTEIN, WHICH INCLUDES AMINO-ACID SEQUENCES OF PROTEIN F OF OUTER MEMBRANE AND EXOTOXIN A PSEUDOMONAS aeruginosa, AND METHOD FOR PRODUCTION OF HYBRID RECOMBINANT PROTEIN, WHICH INCLUDES AMINO-ACID SEQUENCES OF PROTEIN F OF OUTER MEMBRANE AND EXOTOXIN A PSEUDOMONAS aeruginosa</a> <b>[P]</b> . <span> 外国专利：  RU2012139928A </span> <span> . 2014-03-27</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：重组质体DNA pPA-OPRF-ETA，可合成杂种重组蛋白，包括外膜蛋白F的氨基酸序列和铜绿假单胞菌的外毒素，重组大肠杆菌-胰蛋白酶原-蛋白酶。，包括外膜蛋白F和外膜毒素铜绿假单胞菌的氨基酸序列，以及生产杂交重组蛋白的方法，其中包括外膜蛋白F和蛋白质膜外膜蛋白的氨基酸序列 </span> </p> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/patent-detail/06130438427046.html">METHOD FOR MAINTAINING HEALTH, REJUVENATING OR TREATING OR PREVENTING DISEASE BY REPLACING IN VIVO ORGANIC MATTER (CARBOHYDRATE, LIPID, PROTEIN, VITAMINS AND AMINO ACIDS) WITH THAT WITH LOW DEUTERIUM CONCENTRATION AND MANUFACTURING METHOD OF FOOD FOR TAKING ORGANIC MATTER (CARBOHYDRATE, LIPID, PROTEIN, VITAMINS AND AMINO ACIDS) WITH LOW DEUTERIUM CONCENTRATION INTO BODY</a> <b>[P]</b> . <span> 外国专利：  JP2005336146A </span> <span> . 2005-12-08</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：一种方法：用低氘浓度的碳酸氢盐和脂质（制造糖，制造糖的方法）来体内维持，恢复或恢复疾病，方法是用体内有机物质（碳水化合物，脂质，蛋白质，维生素和氨基酸）代替到身体中的低氘浓度的维生素，维生素和氨基酸） </span> </p> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/patent-detail/06130430935992.html">Heat possessed by materials such as heat absorption wavelength bands of amino acids, peptides, proteins, and organic compounds, 2.5 μm to 20 μm area, heat absorption wavelength bands of inorganic metals and semiconductors, and 0.1 μm to 6.5 μm areas Irradiates the wavelength in accordance with the absorption wavelength band with high density, promotes the generation, synthesis, reaction, and decomposition of amino acids, peptides, proteins and organic compounds, and generates inorganic nanoparticles, thin films, and metal crystals. Promote technology development.</a> <b>[P]</b> . <span> 外国专利：  JPWO2007052778A1 </span> <span> . 2009-04-30</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：氨基酸，肽，蛋白质和有机化合物的吸热波长带，2.5μm至20μm面积，无机金属和半导体的吸热波长带以及0.1μm至6.5μm面积等材料所具有的热量根据高密度的吸收波长带，促进氨基酸，肽，蛋白质和有机化合物的生成，合成，反应和分解，并生成无机纳米粒子，薄膜和金属晶体。促进技术发展。 </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('0704028161806','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 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