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bldA编码天蓝色链霉菌中唯一有效识别UUA亮氨酸密码的tRNA(Leu)UUA。通过构建阿维链霉菌NRRL8165基因组亚文库,筛选得到含有阿维链霉菌bldA。及其侧翼序列的克隆。利用λRED介导的PCR targeting技术构建了bldA。的基因置换质粒pHL358,将其跨属接合转移进入阿维链霉菌NRRL8165,筛选得到bldA。基因置换菌株TW10。TW10表现为光秃表型,表明bldA。调控阿维链霉菌的形态分化。摇瓶发酵TW10菌株并对发酵产物进行HPLC分析,发现TW10菌株均不合成阿维菌素组分,提示阿维菌素的合成受bldA。调控;考察阿维菌素生物合成基因簇,其中areA3和aveR含有TTA密码,它们的翻译可能受bldA。调控,与实验结果一致。 相似文献
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Natural products continue to provide privileged scaffolds for drug discovery. However, challenges in supply and structure diversification can limit development. Here, we discuss recent (2017–2020) examples of synthetic biology approaches used to address challenges in supply and contribute to structure diversification of selected plant and bacterial natural products. Our examples include plant terpenoids, alkaloids, and lignans and bacterial polyketides, nonribosomal peptides, and ribosomally synthesized and posttranslationally modified peptides. 相似文献
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链霉菌是革兰氏阳性丝状细菌,其次级代谢产物具有抗感染、抗虫、抗肿瘤、免疫调节等生理活性,在医药、食品和农业领域具有重要应用价值。链霉菌的遗传操作技术是发现和改良新次级代谢产物的基础,近年来合成生物学的兴起为链霉菌的研发提供了全新的视角。综述了合成生物学在链霉菌次级产物生物合成基因簇克隆与组装、底盘细胞设计与改造、调节两者适配性方面的应用进展。 相似文献
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《Studies in History and Philosophy of Science Part C: Studies in History and Philosophy of Biological and Biomedical Sciences》2013,44(2):190-198
The principal existing real-world application of synthetic biology is biofuels. Several ‘next generation biofuel’ companies—Synthetic Genomics, Amyris and Joule Unlimited Technologies—claim to be using synthetic biology to make biofuels. The irony of this is that highly advanced science and engineering serves the very mundane and familiar realm of transport. Despite their rather prosaic nature, biofuels could offer an interesting way to highlight the novelty of synthetic biology from several angles at once. Drawing on the French philosopher of technology and biology Gilbert Simondon, we can understand biofuels as technical objects whose genesis involves processes of concretisation that negotiate between heterogeneous geographical, biological, technical, scientific and commercial realities. Simondon’s notion of technicity, the degree of concretisation of a technical object, usefully conceptualises this relationality. Viewed in terms of technicity, we might understand better how technical entities, elements, and ensembles are coming into being in the name of synthetic biology. The broader argument here is that when we seek to identify the newness of disciplines, their newness might be less epistemic and more logistic. 相似文献
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《Studies in History and Philosophy of Science Part C: Studies in History and Philosophy of Biological and Biomedical Sciences》2013,44(2):208-216
Synthetic biology research is often described in terms of programming cells through the introduction of synthetic genes. Genetic material is seemingly attributed with a high level of causal responsibility. We discuss genetic causation in synthetic biology and distinguish three gene concepts differing in their assumptions of genetic control. We argue that synthetic biology generally employs a difference-making approach to establishing genetic causes, and that this approach does not commit to a specific notion of genetic program or genetic control. Still, we suggest that a strong program concept of genetic material can be used as a successful heuristic in certain areas of synthetic biology. Its application requires control of causal context, and may stand in need of a modular decomposition of the target system. We relate different modularity concepts to the discussion of genetic causation and point to possible advantages of and important limitations to seeking modularity in synthetic biology systems. 相似文献
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目的:对阿佛曼链霉菌采用新的诱变手段,以获得稳定高产的优良菌株。方法:采用重离子束辐照阿佛曼链霉菌,研究了0.25Gy、0.5Gy、3Gy、5Gy、10Gy和15Gy剂量的12C+粒子束辐照阿佛曼链霉菌菌株后,菌落特性的变化及对菌株产素能力的影响。结果:重离子辐照阿佛曼链霉菌后,在其各个辐照剂量区都存在变异菌株,诱变后阿佛曼链霉菌的菌落形态多样,小山状,火山口状、彗星尾状、车轮状、边缘放射状等;菌落大小不一,有的直径达4~5mm,有的小如针尖状。效价提高到7298μg/mL,获得了高产菌株。结论:重离子束辐照阿佛曼链霉菌菌株后,阿佛曼链霉菌的产素能力显著提高,可得到高产的菌株。 相似文献
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Jenny Rooke 《Systems and synthetic biology》2013,7(3):67-72
Synthetic biology has the potential to contribute breakthrough innovations to the pursuit of new global health solutions. Wishing to harness the emerging tools of synthetic biology for the goals of global health, in 2011 the Bill & Melinda Gates Foundation put out a call for grant applications to “Apply Synthetic Biology to Global Health Challenges” under its “Grand Challenges Explorations” program. A highly diverse pool of over 700 applications was received. Proposed applications of synthetic biology to global health needs included interventions such as therapeutics, vaccines, and diagnostics, as well as strategies for biomanufacturing, and the design of tools and platforms that could further global health research. 相似文献
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阿维菌素B1a组分高产菌株的定向选育 总被引:2,自引:0,他引:2
以阿维链霉菌(Streptomyces avermitilis)1-17为出发菌株,分别使用紫外线及亚硝基胍并结合L-异亮氨酸诱导手段进行诱变处理,得到AVMB1a组分摇瓶发酵水平较出发菌株提高12.86%的突变株3-6.传代实验表明该菌株的高产性能稳定.结果表明,采用UV、NTG诱变结合L-Ile诱导的手段可以获得B1a组分显著提高的菌株. 相似文献
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The full potential of polyketide discovery has yet to be reached owing to a lack of suitable technologies and knowledge required to advance engineering of polyketide biosynthesis. Recent investigations on the discovery, enhancement, and non-natural use of these biosynthetic gene clusters via computational biology, metabolic engineering, structural biology, and enzymology-guided approaches have facilitated improved access to designer polyketides. Here, we discuss recent successes in gene cluster discovery, host strain engineering, precursor-directed biosynthesis, combinatorial biosynthesis, polyketide tailoring, and high-throughput synthetic biology, as well as challenges and outlooks for rapidly generating useful target polyketides. 相似文献
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Biological systems are inherently noisy. Predicting the outcome of a perturbation is extremely challenging. Traditional reductionist
approach of describing properties of parts, vis-a-vis higher level behaviour has led to enormous understanding of fundamental
molecular level biology. This approach typically consists of converting genes into junk (knock-down) and garbage (knock-out)
and observe how a system responds. To enable broader understanding of biological dynamics, an integrated computational and
experimental strategy was formally proposed in mid 1990s leading to the re-emergence of Systems Biology. However, soon it
became clear that natural systems were far more complex than expected. A new strategy to address biological complexity was
proposed at MIT (Massachusetts Institute of Technology) in June 2004, when the first meeting of synthetic biology was held.
Though the term ‘synthetic biology’ was proposed during 1970s (Szybalski in Control of gene expression, Plenum Press, New
York, 1974), the usage of the original concept found an experimental proof in 2000 with the demonstration of a three-gene circuit called
repressilator (Elowitz and Leibler in Nature, 403:335–338, 2000). This encouraged people to think of forward engineering biology from a set of well described parts. 相似文献
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Organization of biosynthetic gene cluster for avermectin in Streptomyces avermitilis: analysis of enzymatic domains in four polyketide synthases 总被引:4,自引:0,他引:4
The analysis of the incorporation of 13C-labeled precursors into avermectins indicates that the avermectin aglycons are synthesized by head-to-tail condensation
of various acyl groups, which is similar to the biosynthesis of other polyketides. Polyketide synthases (PKS) use the appropriate
CoA ester as a primer and add acetate units from malonyl-CoA and propionate units from methylmalonyl-CoA to assemble the polyketides.
Avermectin aglycons are formed by addition to the starter unit (2-methylbutyrate or isobutyrate) of 12 acyl condensations
in the order P–A–A–A–A–P–P–A–P–A–P–A (P, propionyl; A, acetyl). Within the 90-kb gene cluster for avermectin biosynthesis,
the central 65-kb segment was found to be required for aglycon biosynthesis by phenotypic analysis of strains containing deletion
or insertion mutations in this region. A complete sequence analysis of the 65-kb segment indicated that this segment encodes
avermectin PKS. The avermectin PKS genes are organized into two converging blocks of ORFs. From the results of sequencing
analysis, a feature of the two regions, aveA1/aveA2 and avea3/aveA4, is that they encode four kinds of large multifunctional polypeptides containing 55 domains which possess putative fatty
acid synthase-like activities. The avermectin PKS (AVES 1–4) appear to contain two, three, or four modules. AVES 1 and 2 contain
two and four modules, respectively, whereas AVES 3 and AVES 4 each contains three modules. The 12 modules correspond to the
12 cycles required for synthesis of the avermectin aglycon. Journal of Industrial Microbiology & Biotechnology (2001) 27, 170–176.
Received 21 September 1999/ Accepted in revised form 14 September 2000 相似文献
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【目的】考察除虫链霉菌基因组中其它聚酮合成酶类(Polyketide synthase,PKS)抗生素生物合成基因簇的敲除突变对于阿维菌素产量的影响。【方法】构建了11个PKS基因簇的打靶Cosmid和质粒载体,导入除虫链霉菌中筛选突变株。【结果】在工业菌株MMR630中成功敲除了10个PKS基因簇。发酵结果显示7个PKS基因簇敲除突变株中阿维菌素的产量均有不同程度的提高,而2个突变株不能产生阿维菌素。然而,在3个连续敲除2个PKS基因簇的突变株中阿维菌素产量没有能够超过单个PKS敲除突变株的提升幅度。【结论】除虫链霉菌基因组的一些PKS基因簇的敲除可以提高阿维菌素的产量,同时暗示同一类次生代谢产物的代谢流之间存在复杂的相互作用关系。 相似文献
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The non-conventional oleaginous yeast Yarrowia lipolytica shows great industrial promise. It naturally produces certain compounds of interest but can also artificially generate non-native metabolites, thanks to an engineering process made possible by the significant expansion of a dedicated genetic toolbox. In this review, we present recently developed synthetic biology tools that facilitate the manipulation of Y. lipolytica, including 1) DNA assembly techniques, 2) DNA parts for constructing expression cassettes, 3) genome-editing techniques, and 4) computational tools. 相似文献
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《Studies in History and Philosophy of Science Part C: Studies in History and Philosophy of Biological and Biomedical Sciences》2013,44(2):199-207
The parts-based engineering approach in synthetic biology aims to create pre-characterised biological parts that can be used for the rational design of novel functional systems. Given the context-sensitivity of biological entities, a key question synthetic biologists have to address is what properties these parts should have so that they give a predictable output even when they are used in different contexts. In the first part of this paper I will analyse some of the answers that synthetic biologists have given to this question and claim that the focus of these answers on parts and their properties does not allow us to tackle the problem of context-sensitivity. In the second part of the paper, I will argue that we might have to abandon the notions of parts and their properties in order to understand how independence in biology could be achieved. Using Robert Cummins’ account of functional analysis, I will then develop the notion of a capacity and its condition space and show how these notions can help to tackle the problem of context-sensitivity in biology. 相似文献
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随着基因回路规模的扩大,和应用范围的拓展,传统的合成基因回路的设计思路面临着新的挑战。新合成基因回路构建的试验周期长,试错成本大,单纯依靠经验进行设计构建,难以迅速得到满意的结果。iGEM中软件设计比赛旨在帮助合成生物学家,更高效地完成基因回路的设计与预测。为了更好地研究iGEM软件的设计与研究方向,寻找新的设计思路和理念,综述了最近几年iGEM软件队的项目,仔细总结了每一个项目的背景、目的,设计和应用。通过对比和总结,发现这几年的iGEM软件项目从功能上可以分为以下四类:①辅助设计;②资料共享;③合作交流;④数据分析。该综述可以为今后iGEM软件设计提供思考方向,也为合成生物学的发展提供新的思路。 相似文献
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Synthetic Biology is a rapidly growing interdisciplinary field that is primarily built upon foundational advances in molecular biology combined with engineering design principles such as modularity and interoperability. The field considers living systems as programmable at the genetic level and has been defined by the development of new platform technologies and methodological advances. A key concept driving the field is the Design-Build-Test-Learn cycle which provides a systematic framework for building new biological systems. One major application area for synthetic biology is biosynthetic pathway engineering that requires the modular assembly of different genetic regulatory elements and biosynthetic enzymes. In this review we provide an overview of modular DNA assembly and describe and compare the plethora of in vitro and in vivo assembly methods for combinatorial pathway engineering. Considerations for part design and methods for enzyme balancing are also presented, and we briefly discuss alternatives to intracellular pathway assembly including microbial consortia and cell-free systems for biosynthesis. Finally, we describe computational tools and automation for pathway design and assembly and argue that a deeper understanding of the many different variables of genetic design, pathway regulation and cellular metabolism will allow more predictive pathway design and engineering. 相似文献