Anna B. Matuszyńska, O. Ebenhöh, Matias D Zurbriggen, Daniel C Ducat, Ilka M. Axmann
Synthetic biology conceptualises biological complexity as a network of biological parts, devices and systems with predetermined functionalities, and has had a revolutionary impact on fundamental and applied research. With the unprecedented ability to synthesise and transfer any DNA and RNA across organisms, the scope of synthetic biology is expanding and being recreated in previously unimaginable ways. The field has matured to a level where highly complex networks, such as artificial communities of synthetic organisms can be constructed. In parallel, computational biology became an integral part of biological studies, with computational models aiding the unravelling of the escalating complexity and emerging properties of biological phenomena. However, there is still a vast untapped potential for the complete integration of modelling into the synthetic design process, presenting exciting opportunities for scientific advancements. Here, we first highlight the most recent advances in computer-aided design of microbial communities. Next, we propose that such a design can benefit from an organism-free modular modelling approach that places its emphasis on modules of organismal function towards the design of multi-species communities. We argue for a shift in perspective from single organism-centred approaches to emphasising the functional contributions of organisms within the community. By assembling synthetic biological systems using modular computational models with mathematical descriptions of parts and circuits, we can tailor organisms to fulfil specific functional roles within the community. This approach aligns with synthetic biology strategies and presents exciting possibilities for the design of artificial communities.
合成生物学将生物复杂性概念化为具有预定功能的生物部件、装置和系统网络，对基础研究和应用研究产生了革命性的影响。由于合成和跨生物体转移任何 DNA 和 RNA 的能力前所未有，合成生物学的范围不断扩大，并以以前无法想象的方式进行再创造。该领域已经发展到可以构建高度复杂网络的水平，例如合成生物的人工群落。与此同时，计算生物学也成为生物学研究不可或缺的一部分，计算模型有助于揭示生物现象不断升级的复杂性和新特性。然而，将建模完全融入合成设计过程仍有巨大的潜力尚未开发，这为科学进步带来了令人兴奋的机遇。在此，我们首先重点介绍微生物群落计算机辅助设计的最新进展。接下来，我们提出，这种设计可以受益于无生物模块建模方法，这种方法将重点放在生物功能模块上，从而设计出多物种群落。我们主张转变视角，从以单一生物为中心的方法转向强调群落内生物的功能贡献。通过使用具有部件和电路数学描述的模块化计算模型组装合成生物系统，我们可以定制生物体，使其在群落中发挥特定的功能作用。这种方法符合合成生物学战略，为设计人工群落提供了令人兴奋的可能性。
{"title":"A new era of Synthetic Biology - microbial community design","authors":"Anna B. Matuszyńska, O. Ebenhöh, Matias D Zurbriggen, Daniel C Ducat, Ilka M. Axmann","doi":"10.1093/synbio/ysae011","DOIUrl":"https://doi.org/10.1093/synbio/ysae011","url":null,"abstract":"\u0000 Synthetic biology conceptualises biological complexity as a network of biological parts, devices and systems with predetermined functionalities, and has had a revolutionary impact on fundamental and applied research. With the unprecedented ability to synthesise and transfer any DNA and RNA across organisms, the scope of synthetic biology is expanding and being recreated in previously unimaginable ways. The field has matured to a level where highly complex networks, such as artificial communities of synthetic organisms can be constructed. In parallel, computational biology became an integral part of biological studies, with computational models aiding the unravelling of the escalating complexity and emerging properties of biological phenomena. However, there is still a vast untapped potential for the complete integration of modelling into the synthetic design process, presenting exciting opportunities for scientific advancements. Here, we first highlight the most recent advances in computer-aided design of microbial communities. Next, we propose that such a design can benefit from an organism-free modular modelling approach that places its emphasis on modules of organismal function towards the design of multi-species communities. We argue for a shift in perspective from single organism-centred approaches to emphasising the functional contributions of organisms within the community. By assembling synthetic biological systems using modular computational models with mathematical descriptions of parts and circuits, we can tailor organisms to fulfil specific functional roles within the community. This approach aligns with synthetic biology strategies and presents exciting possibilities for the design of artificial communities.","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141643800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The paper addresses the application of engineering biology strategy and techniques to the automation of laboratory workflow - primarily in the context of biofoundries and biodesign applications based on the Design, Build, Test and Learn paradigm. The trend towards greater automation comes with its own set of challenges. On the one hand, automation is associated with higher throughput and with higher replicability. On the other hand, implementation of an automated workflow requires an instruction set that is far more extensive than for a manual workflow. Automated tasks must also be conducted in the order specified in the workflow, with the right logic, utilising suitable biofoundry resources, and at scale - whilst simultaneously collecting measurements and associated data. The paper describes an approach to automated workflow that is being trialled at the London Biofoundry at SynbiCITE. The solution represents workflows with directed graphs, uses orchestrators for their execution and relies on existing standards. The approach is highly flexible and applies to not only workflow automation in single locations but also distributed workflows (e.g for biomanufacturing). The final section presents an overview of the implementation - using the simple example of an assay based on a dilution, measurement and data analysis workflow.
{"title":"An Engineering Biology Approach to Automated Workflow and BioDesign","authors":"Alexis Casas, Matthieu Bultelle, Richard Kitney","doi":"10.1093/synbio/ysae009","DOIUrl":"https://doi.org/10.1093/synbio/ysae009","url":null,"abstract":"\u0000 The paper addresses the application of engineering biology strategy and techniques to the automation of laboratory workflow - primarily in the context of biofoundries and biodesign applications based on the Design, Build, Test and Learn paradigm. The trend towards greater automation comes with its own set of challenges. On the one hand, automation is associated with higher throughput and with higher replicability. On the other hand, implementation of an automated workflow requires an instruction set that is far more extensive than for a manual workflow. Automated tasks must also be conducted in the order specified in the workflow, with the right logic, utilising suitable biofoundry resources, and at scale - whilst simultaneously collecting measurements and associated data.\u0000 The paper describes an approach to automated workflow that is being trialled at the London Biofoundry at SynbiCITE. The solution represents workflows with directed graphs, uses orchestrators for their execution and relies on existing standards. The approach is highly flexible and applies to not only workflow automation in single locations but also distributed workflows (e.g for biomanufacturing).\u0000 The final section presents an overview of the implementation - using the simple example of an assay based on a dilution, measurement and data analysis workflow.","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141337572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ludo L J Schoenmakers, Max J den Uijl, Jelle Postma, Tim A P van den Akker, Wilhelm T S Huck, Arnold J. M. Driessen
Giant unilamellar vesicles (GUVs) provide a powerful model compartment for synthetic cells. However, a key challenge is the incorporation of membrane proteins that allow for transport, energy transduction, compartment growth and division. Here, we have successfully incorporated the membrane protein complex SecYEG – the key bacterial translocase that is essential for the incorporation of newly synthesized membrane proteins – in GUVs. Our method consists of fusion of small unilamellar vesicles (SUVs) containing reconstituted SecYEG into GUVs, thereby forming SecGUVs. These are suitable for large scale experiments while maintaining a high protein:lipid ratio. We demonstrate that incorporation of SecYEG into GUVs does not inhibit its translocation efficiency. Robust membrane protein functionalized proteo-GUVs are promising and flexible compartments for use in the formation and growth of synthetic cells.
{"title":"SecYEG-mediated Translocation in a Model Synthetic Cell","authors":"Ludo L J Schoenmakers, Max J den Uijl, Jelle Postma, Tim A P van den Akker, Wilhelm T S Huck, Arnold J. M. Driessen","doi":"10.1093/synbio/ysae007","DOIUrl":"https://doi.org/10.1093/synbio/ysae007","url":null,"abstract":"\u0000 Giant unilamellar vesicles (GUVs) provide a powerful model compartment for synthetic cells. However, a key challenge is the incorporation of membrane proteins that allow for transport, energy transduction, compartment growth and division. Here, we have successfully incorporated the membrane protein complex SecYEG – the key bacterial translocase that is essential for the incorporation of newly synthesized membrane proteins – in GUVs. Our method consists of fusion of small unilamellar vesicles (SUVs) containing reconstituted SecYEG into GUVs, thereby forming SecGUVs. These are suitable for large scale experiments while maintaining a high protein:lipid ratio. We demonstrate that incorporation of SecYEG into GUVs does not inhibit its translocation efficiency. Robust membrane protein functionalized proteo-GUVs are promising and flexible compartments for use in the formation and growth of synthetic cells.","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140992187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A new Editor-in-chief for Synthetic Biology","authors":"Sonja Billerbeck","doi":"10.1093/synbio/ysae006","DOIUrl":"https://doi.org/10.1093/synbio/ysae006","url":null,"abstract":"","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140697028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Writing the Dark Matter of the Human Genome into Mice to better replicate human disease.","authors":"David M. Truong","doi":"10.1093/synbio/ysae003","DOIUrl":"https://doi.org/10.1093/synbio/ysae003","url":null,"abstract":"","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139619116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"On the path to generate electricity from wastewater through genetic engineering of Escherichia coli","authors":"C. Cialek","doi":"10.1093/synbio/ysae002","DOIUrl":"https://doi.org/10.1093/synbio/ysae002","url":null,"abstract":"","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139623731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alexander Ditzel, Fanglong Zhao, Xue Gao, George N Phillips
Natural products are a valuable source of pharmaceuticals, providing a majority of the small molecule drugs in use today. However, their production through organic synthesis or in heterologous hosts can be difficult and time-consuming. Therefore, to allow for easier screening and production of natural products, we demonstrated the use of a cell-free protein synthesis (CFPS) system to partially assemble natural products in vitro using SAM-dependent methyltransferase enzyme reactions. The tea caffeine synthase TCS1 was utilized to synthesize caffeine within a CFPS system. Cell-free systems also provide the benefit of allowing the use of substrates that would normally be toxic in a cellular environment to synthesize novel products. However, TCS1 is unable to utilize a compound like AdoEt as a cofactor to create ethylated caffeine analogs. The automation and reduced metabolic engineering requirements of CFPS systems, in combination with other synthesis methods, may enable the more efficient generation of new compounds.
{"title":"Utilizing a Cell-free Protein Synthesis Platform for the Biosynthesis of a Natural Product, Caffeine","authors":"Alexander Ditzel, Fanglong Zhao, Xue Gao, George N Phillips","doi":"10.1093/synbio/ysad017","DOIUrl":"https://doi.org/10.1093/synbio/ysad017","url":null,"abstract":"\u0000 Natural products are a valuable source of pharmaceuticals, providing a majority of the small molecule drugs in use today. However, their production through organic synthesis or in heterologous hosts can be difficult and time-consuming. Therefore, to allow for easier screening and production of natural products, we demonstrated the use of a cell-free protein synthesis (CFPS) system to partially assemble natural products in vitro using SAM-dependent methyltransferase enzyme reactions. The tea caffeine synthase TCS1 was utilized to synthesize caffeine within a CFPS system. Cell-free systems also provide the benefit of allowing the use of substrates that would normally be toxic in a cellular environment to synthesize novel products. However, TCS1 is unable to utilize a compound like AdoEt as a cofactor to create ethylated caffeine analogs. The automation and reduced metabolic engineering requirements of CFPS systems, in combination with other synthesis methods, may enable the more efficient generation of new compounds.","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138945344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Matthew W Lux, Elizabeth A Strychalski, Gary J Vora
Abstract Reproducibility has been identified as an outstanding challenge in science, and the field of synthetic biology is no exception. Meeting this challenge is critical to allow the transformative technological capabilities emerging from this field to reach their full potential to benefit the society. We discuss the current state of reproducibility in synthetic biology and how improvements can address some of the central shortcomings in the field. We argue that the successful adoption of reproducibility as a routine aspect of research and development requires commitment spanning researchers and relevant institutions via education, incentivization and investment in related infrastructure. The urgency of this topic pervades synthetic biology as it strives to advance fundamental insights and unlock new capabilities for safe, secure and scalable applications of biotechnology. Graphical Abstract
{"title":"Advancing reproducibility can ease the ‘hard truths’ of synthetic biology","authors":"Matthew W Lux, Elizabeth A Strychalski, Gary J Vora","doi":"10.1093/synbio/ysad014","DOIUrl":"https://doi.org/10.1093/synbio/ysad014","url":null,"abstract":"Abstract Reproducibility has been identified as an outstanding challenge in science, and the field of synthetic biology is no exception. Meeting this challenge is critical to allow the transformative technological capabilities emerging from this field to reach their full potential to benefit the society. We discuss the current state of reproducibility in synthetic biology and how improvements can address some of the central shortcomings in the field. We argue that the successful adoption of reproducibility as a routine aspect of research and development requires commitment spanning researchers and relevant institutions via education, incentivization and investment in related infrastructure. The urgency of this topic pervades synthetic biology as it strives to advance fundamental insights and unlock new capabilities for safe, secure and scalable applications of biotechnology. Graphical Abstract","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135611266","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-10-06eCollection Date: 2022-01-01DOI: 10.1093/synbio/ysac018
Robert P Goldman, Robert Moseley, Nicholas Roehner, Breschine Cummins, Justin D Vrana, Katie J Clowers, Daniel Bryce, Jacob Beal, Matthew DeHaven, Joshua Nowak, Trissha Higa, Vanessa Biggers, Peter Lee, Jeremy P Hunt, Lorraine Mosqueda, Steven B Haase, Mark Weston, George Zheng, Anastasia Deckard, Shweta Gopaulakrishnan, Joseph F Stubbs, Niall I Gaffney, Matthew W Vaughn, Narendra Maheshri, Ekaterina Mikhalev, Bryan Bartley, Richard Markeloff, Tom Mitchell, Tramy Nguyen, Daniel Sumorok, Nicholas Walczak, Chris Myers, Zach Zundel, Benjamin Hatch, James Scholz, John Colonna-Romano
We describe an experimental campaign that replicated the performance assessment of logic gates engineered into cells of Saccharomyces cerevisiae by Gander et al. Our experimental campaign used a novel high-throughput experimentation framework developed under Defense Advanced Research Projects Agency's Synergistic Discovery and Design program: a remote robotic lab at Strateos executed a parameterized experimental protocol. Using this protocol and robotic execution, we generated two orders of magnitude more flow cytometry data than the original experiments. We discuss our results, which largely, but not completely, agree with the original report and make some remarks about lessons learned. Graphical Abstract.
{"title":"Highly-automated, high-throughput replication of yeast-based logic circuit design assessments.","authors":"Robert P Goldman, Robert Moseley, Nicholas Roehner, Breschine Cummins, Justin D Vrana, Katie J Clowers, Daniel Bryce, Jacob Beal, Matthew DeHaven, Joshua Nowak, Trissha Higa, Vanessa Biggers, Peter Lee, Jeremy P Hunt, Lorraine Mosqueda, Steven B Haase, Mark Weston, George Zheng, Anastasia Deckard, Shweta Gopaulakrishnan, Joseph F Stubbs, Niall I Gaffney, Matthew W Vaughn, Narendra Maheshri, Ekaterina Mikhalev, Bryan Bartley, Richard Markeloff, Tom Mitchell, Tramy Nguyen, Daniel Sumorok, Nicholas Walczak, Chris Myers, Zach Zundel, Benjamin Hatch, James Scholz, John Colonna-Romano","doi":"10.1093/synbio/ysac018","DOIUrl":"10.1093/synbio/ysac018","url":null,"abstract":"<p><p>We describe an experimental campaign that replicated the performance assessment of logic gates engineered into cells of <i>Saccharomyces cerevisiae</i> by Gander <i>et al.</i> Our experimental campaign used a novel high-throughput experimentation framework developed under Defense Advanced Research Projects Agency's Synergistic Discovery and Design program: a remote robotic lab at Strateos executed a parameterized experimental protocol. Using this protocol and robotic execution, we generated two orders of magnitude more flow cytometry data than the original experiments. We discuss our results, which largely, but not completely, agree with the original report and make some remarks about lessons learned. <b>Graphical Abstract</b>.</p>","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2022-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9583850/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87759237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Recombineering is an important tool in gene editing, enabling fast, precise and highly specific in vivo modification of microbial genomes. Oligonucleotide-mediated recombineering via the in vivo production of single-stranded DNA can overcome the limitations of traditional recombineering methods that rely on the exogenous delivery of editing templates. By modifying a previously reported plasmid-based system for fully in vivo single-stranded DNA recombineering, we demonstrate iterative editing of independent loci by utilizing a temperature-sensitive origin of replication for easy curing of the editing plasmid from recombinant cells. Optimization of the promoters driving the expression of the system’s functional components, combined with targeted counterselection against unedited cells with Cas9 nuclease, enabled editing efficiencies of 90–100%. The addition of a dominant-negative mutL allele to the system allowed single-nucleotide edits that were otherwise unachievable due to mismatch repair. Finally, we tested alternative recombinases and found that efficiency significantly increased for some targets. Requiring only a single cloning step for retargeting, our system provides an easy-to-use method for rapid, efficient construction of desired mutants. Graphical Abstract
{"title":"Efficient and iterative retron-mediated in vivo recombineering in Escherichia coli","authors":"A. Ellington, Christopher R. Reisch","doi":"10.1093/synbio/ysac007","DOIUrl":"https://doi.org/10.1093/synbio/ysac007","url":null,"abstract":"Abstract Recombineering is an important tool in gene editing, enabling fast, precise and highly specific in vivo modification of microbial genomes. Oligonucleotide-mediated recombineering via the in vivo production of single-stranded DNA can overcome the limitations of traditional recombineering methods that rely on the exogenous delivery of editing templates. By modifying a previously reported plasmid-based system for fully in vivo single-stranded DNA recombineering, we demonstrate iterative editing of independent loci by utilizing a temperature-sensitive origin of replication for easy curing of the editing plasmid from recombinant cells. Optimization of the promoters driving the expression of the system’s functional components, combined with targeted counterselection against unedited cells with Cas9 nuclease, enabled editing efficiencies of 90–100%. The addition of a dominant-negative mutL allele to the system allowed single-nucleotide edits that were otherwise unachievable due to mismatch repair. Finally, we tested alternative recombinases and found that efficiency significantly increased for some targets. Requiring only a single cloning step for retargeting, our system provides an easy-to-use method for rapid, efficient construction of desired mutants. Graphical Abstract","PeriodicalId":22158,"journal":{"name":"Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2022-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74123254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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