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  • 文章类型: Journal Article
    Przondovirus属中的噬菌体(噬菌体)是属于Studiervirinae亚科的T7样podovirus,在自拟病毒科中,并且具有高度保守的基因组组织。这些噬菌体的基因组大小从37到42kb,编码50-60个基因,其特征在于存在线性染色体侧翼的直接末端重复(DTR)。这些DTR通常在短只读和混合组装期间被删除。此外,长只读组件通常会出现测序和/或组装错误,需要额外的管理。这里,我们介绍了十种针对克雷伯菌属的新型przondovirus的分离和表征。我们描述了HYPPA,混合和聚波兰噬菌体组装工作流程,它利用长读取组件与短读取测序相结合来解析噬菌体DTR并纠正错误,否定了费力的引物行走和Sanger测序验证的需要。我们的组装工作流程利用牛津纳米孔技术进行长读取测序,使其成为目前更相关的长读数测序技术,和IlluminaDNA准备进行短读测序,代表全球最常用的技术。我们的数据证明了在发表之前仔细管理噬菌体组装的重要性,在将它们用于比较基因组学之前。
    Bacteriophages (phages) within the genus Przondovirus are T7-like podoviruses belonging to the subfamily Studiervirinae, within the family Autographiviridae, and have a highly conserved genome organisation. The genomes of these phages range from 37 to 42 kb in size, encode 50-60 genes and are characterised by the presence of direct terminal repeats (DTRs) flanking the linear chromosome. These DTRs are often deleted during short-read-only and hybrid assemblies. Moreover, long-read-only assemblies are often littered with sequencing and/or assembly errors and require additional curation. Here, we present the isolation and characterisation of ten novel przondoviruses targeting Klebsiella spp. We describe HYPPA, a HYbrid and Poly-polish Phage Assembly workflow, which utilises long-read assemblies in combination with short-read sequencing to resolve phage DTRs and correcting errors, negating the need for laborious primer walking and Sanger sequencing validation. Our assembly workflow utilised Oxford Nanopore Technologies for long-read sequencing for its accessibility, making it the more relevant long-read sequencing technology at this time, and Illumina DNA Prep for short-read sequencing, representing the most commonly used technologies globally. Our data demonstrate the importance of careful curation of phage assemblies before publication, and prior to using them for comparative genomics.
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  • 文章类型: Journal Article
    随着长读测序仪和远程技术的兴起,提供高质量的植物基因组组装不再保留给大型财团。不仅仅是测序技术,而且,计算机算法已经达到了在实验室规模上现在可以在染色体规模上重建组件的地步。当前技术,特别是远程技术,很多,和选择最有前途的一个感兴趣的基因组是至关重要的,以获得最佳结果。在这项研究中,我们对黄萨森的基因组进行了重新测序,芸苔属拉帕简历。Z1,使用牛津纳米孔PromethION测序仪,并使用当前的汇编器组装测序数据。为了重建完整的染色体,我们使用并比较了三种远程脚手架技术,光学测绘,Omni-C,和Pore-C测序文库,由Bionano基因组学商业化,燕尾基因组学,和牛津纳米孔技术,分别,或者三者的组合,以评估每种技术的能力。
    With the rise of long-read sequencers and long-range technologies, delivering high-quality plant genome assemblies is no longer reserved to large consortia. Not only sequencing techniques, but also computer algorithms have reached a point where the reconstruction of assemblies at the chromosome scale is now feasible at the laboratory scale. Current technologies, in particular long-range technologies, are numerous, and selecting the most promising one for the genome of interest is crucial to obtain optimal results. In this study, we resequenced the genome of the yellow sarson, Brassica rapa cv. Z1, using the Oxford Nanopore PromethION sequencer and assembled the sequenced data using current assemblers. To reconstruct complete chromosomes, we used and compared three long-range scaffolding techniques, optical mapping, Omni-C, and Pore-C sequencing libraries, commercialized by Bionano Genomics, Dovetail Genomics, and Oxford Nanopore Technologies, respectively, or a combination of the three, in order to evaluate the capability of each technology.
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  • 文章类型: Journal Article
    具有光热剂的化疗药物的自组装纳米载体被认为是有趣的化学-光热疗法纳米平台。然而,大多数药物和光热剂的水溶性差和相互作用差,以驱动自组装纳米载体的形成,这是共同组装的药物/光热剂用于癌症治疗的瓶颈。这里,我们提出了一种基于光热剂和药物化学修饰的自组装化学-光热疗法纳米平台的通用策略。选择IR-780和喜树碱(CPT)作为研究模型,因为它们是重要的光热剂和抗癌药物,它们都具有如此差的水溶性和强烈的自身分子相互作用,以至于它们不能共组装在一起。IR-780用活性靶向配体乳糖酸(LA)修饰以产生两亲性IR780-LA,而CPT通过二硫键修饰成氧化还原敏感的前药CPT-ss-CPT以实现其组装。可以通过IR780-LA和CPT-ss-CPT的共组装来产生明确定义的纳米颗粒(NP)。IR780-LA/CPT-ss-CPT纳米粒子被证明是一种出色的荧光成像引导,氧化还原响应和增强协同化学光热疗法纳米平台抗肿瘤。具体来说,我们的化学修饰策略提供了一种通用的方法来创建自组装的化学光热疗法纳米平台,解决了共组装药物/光热剂用于癌症治疗的瓶颈。重要性声明:具有光热药物的化学治疗剂的自组装纳米颗粒被认为是有趣的化学-光热疗法纳米平台。然而,大多数药物的溶解性和相互作用都很差,无法形成自组装的纳米颗粒。我们提出了一种通用策略,基于常见药物的化学修饰来创建共组装的化学-光热治疗纳米颗粒。IR-780用活性靶向配体LA修饰,产生两亲性IR780-LA分子,而CPT通过二硫键修饰为氧化还原敏感的前药CPT-ss-CPT。明确定义的IR780-LA/CPT-ss-CPT纳米颗粒通过IR780-LA和CPT-ss-CPT的共组装而产生。纳米粒子被证明是一种优异的荧光成像引导,氧化还原响应,主动靶向化学光热治疗纳米平台抗肿瘤。我们的策略提供了一种从常见药物构建智能化学光热治疗纳米平台的通用方法。
    Self-assembled nanovehicles of chemotherapy drug with photothermal agent are regarded as intriguing chemo-photothermal therapy nanoplatform. However, most of the drugs and photothermal agents have poor water solubility and poor interactions to drive the formation of self-assembled nanovehicles, which is a bottleneck of co-assembled drug/photothermal agent for cancer therapy. Here, we proposed a versatile strategy to create self-assembled chemo-photothermal therapy nanoplatform based on the chemical modification of photothermal agent and drug. The IR-780 and camptothecin (CPT) were chosen as the studied models since they are important photothermal agent and anticancer drug, both of which have such poor water solubility with strong itself molecular interactions that they cannot co-assemble together. IR-780 was modified with an active targeting ligand lactobionic acid (LA) to result in amphiphilic IR780-LA while CPT was modified into redox-sensitive prodrug CPT-ss-CPT through a disulfide linkage to realize its assembly. Well-defined nanoparticles (NPs) could be created through the co-assembling of IR780-LA and CPT-ss-CPT. The IR780-LA/CPT-ss-CPT nanoparticles were demonstrated to be an excellent fluorescence imaging-guided, redox-responsive and enhanced synergistic chemo-photothermal therapy nanoplatform against tumors. Specifically, our chemical modification strategy offers a universal way to create self-assembled chemo-photothermal therapy nanoplatform, which solves the bottleneck of co-assembled drug/photothermal agent for cancer therapy. STATEMENT OF SIGNIFICANCE: Self-assembled nanoparticles of chemotherapeutics with photothermic drugs are regarded as intriguing chemo-photothermal therapy nanoplatform. However, most drugs have too poor solubility and interactions to form into self-assembled nanoparticles. We proposed a versatile strategy to create co-assembled chemo-photothermal therapy nanoparticles based on the chemical modification of common drugs. The IR-780 was modified with an active targeting ligand LA to result in amphiphilic IR780-LA molecules, while CPT was modified into redox-sensitive prodrug CPT-ss-CPT through disulfide linkage. Well-defined IR780-LA/CPT-ss-CPT nanoparticles were created through the co-assembling of IR780-LA and CPT-ss-CPT. The nanoparticles were demonstrated to be an excellent fluorescence imaging-guided, redox-responsive, active targeting chemo-photothermal therapy nanoplatform against tumors. Our strategy offers a versatile way to construct smart chemo-photothermal therapy nanoplatform from common drugs.
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  • 文章类型: Journal Article
    Advances in genomic sequencing technologies in the past decade have revolutionized the field of genomics, resulting in faster and less expensive sequencing. Holding back the potential for innovation, however, is a widespread lack of understanding of genomics and sequencing by the general public. In an attempt to remedy this problem, this paper presents an introduction to the fields of genomics, bioinformatics, and proteomics using the blueberry genome as a model case study of the plant genomics field. The blueberry (Vaccinium sect. Cyanococcus) is often cited as a \"super food\" in the media due to its nutritional benefits and global economic importance. There have been a number of related genomic publications in the past 20 years; however, a completed genome and a full analysis into the health-related pathways are still needed. As exemplified by this blueberry case study, there are opportunities for future genomic research into numerous beneficial plant species. The solid background presented in this paper provides future researchers the foundation to explore these uncharted areas.
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