Genetic Techniques

遗传技术
  • 文章类型: Journal Article
    端粒位于染色体的末端,具有具有保护基因的独特结构的特定序列。它们具有保护染色体末端免受融合事件并确保染色体稳定性的加帽结构。端粒在细胞分裂的每个周期中长度缩短。当这个长度达到某个阈值时,它会导致基因组不稳定,因此牵涉到各种疾病,包括癌症和神经退行性疾病。正在探索端粒作为衰老和年龄相关疾病的生物标志物的可能性,其意义仍在研究中。这是因为有丝分裂后的细胞,它们是不经历有丝分裂的成熟细胞,不要因年龄而经历端粒缩短。相反,其他原因,例如,暴露于氧化应激,会直接损伤端粒,导致基因组不稳定。尽管如此,已经建立了一个普遍的共识,即测量端粒长度提供了有价值的见解,并为分析基因表达和表观遗传数据奠定了重要的基础。已经开发了许多方法来精确测量端粒长度。在这次审查中,我们总结了评估端粒长度的各种方法及其优点和局限性。
    Telomeres are located at the ends of chromosomes and have specific sequences with a distinctive structure that safeguards genes. They possess capping structures that protect chromosome ends from fusion events and ensure chromosome stability. Telomeres shorten in length during each cycle of cell division. When this length reaches a certain threshold, it can lead to genomic instability, thus being implicated in various diseases, including cancer and neurodegenerative disorders. The possibility of telomeres serving as a biomarker for aging and age-related disease is being explored, and their significance is still under study. This is because post-mitotic cells, which are mature cells that do not undergo mitosis, do not experience telomere shortening due to age. Instead, other causes, for example, exposure to oxidative stress, can directly damage the telomeres, causing genomic instability. Nonetheless, a general agreement has been established that measuring telomere length offers valuable insights and forms a crucial foundation for analyzing gene expression and epigenetic data. Numerous approaches have been developed to accurately measure telomere lengths. In this review, we summarize various methods and their advantages and limitations for assessing telomere length.
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  • 文章类型: Journal Article
    与野生动物有关的犯罪是全球第二大最普遍的违法行为。这种非法贸易包括狩猎,繁殖和贩运。除了减少许多物种及其栖息地和生态系统,阻碍依赖它们的当地社区的经济发展,破坏法治和资助恐怖主义,病原体的各种跨物种传播(人畜共患病),包括COVID-19,可以归因于野生动物犯罪。野生动物取证应用跨学科的科学分析,以支持执法调查野生动物犯罪。它的主要目标是确定有问题的分类物种,确定是否犯罪,将嫌疑人与犯罪联系起来,并支持对肇事者的定罪和起诉。本文回顾了野生动物犯罪及其影响,野生动物法医学调查,常见形式的野生动物生物学证据,包括DNA,野生动物DNA技术和野生动物法医遗传学中的挑战。文章还综述了短串联重复序列(STR)和线粒体DNA(mtDNA)标记等遗传标记的贡献,它提供了证明遗传数据,代表了解决野生动物犯罪的大部分DNA证据。这篇综述概述了野生动物DNA数据库,这对于搜索和匹配法医DNA谱和序列以及确定法医DNA谱和序列在特定人群或地理区域中的频繁程度至关重要。因此,这篇综述将深入分析野生动物法医遗传学的现状,这将是野生动物和保护生物学家普遍感兴趣的,执法人员,和学者对使用动物法医DNA方法打击针对野生动物的犯罪感兴趣。
    Wildlife-related crimes are the second most prevalent lawbreaking offense globally. This illicit trade encompasses hunting, breeding and trafficking. Besides diminishing many species and their habitats and ecosystems, hindering the economic development of local communities that depend on them, undermining the rule of law and financing terrorism, various cross-species transmissions (zoonoses) of pathogens, including COVID-19, can be attributed to wildlife crimes. Wildlife forensics applies interdisciplinary scientific analyses to support law enforcement in investigating wildlife crimes. Its main objectives are to identify the taxonomic species in question, determine if a crime has been committed, link a suspect to the crime and support the conviction and prosecution of the perpetrator. This article reviews wildlife crime and its implications, wildlife forensic science investigation, common forms of wildlife biological evidence, including DNA, wildlife DNA techniques and challenges in wildlife forensic genetics. The article also reviews the contributions of genetic markers such as short tandem repeat (STR) and mitochondrial DNA (mtDNA) markers, which provide the probative genetic data representing the bulk of DNA evidence for solving wildlife crime. This review provides an overview of wildlife DNA databases, which are critical for searching and matching forensic DNA profiles and sequences and establishing how frequent forensic DNA profiles and sequences are in a particular population or geographic region. As such, this review will contain an in-depth analysis of the current status of wildlife forensic genetics, and it will be of general interest to wildlife and conservation biologists, law enforcement officers, and academics interested in combating crimes against wildlife using animal forensic DNA methods.
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  • 文章类型: Journal Article
    阿卡波糖作为α-葡萄糖苷酶抑制剂广泛用于治疗Ⅱ型糖尿病。放线飞机sp.用于工业生产阿卡波糖。作为次级代谢产物,阿卡波糖的生物合成相当复杂。除了阿卡波糖,在放线菌属的培养液中也积累了一些阿卡波糖结构类似物。,很难删除。由于缺乏对阿卡波糖及其结构类似物的生物合成和调节机制的系统理解,很难消除或减少结构类似物的生物合成。最近,组学技术和分子生物学的进步促进了放线菌中阿卡波糖及其结构类似物的生物合成和调节机制的研究。.通过生物信息学分析,对阿卡波糖及其结构类似物的生物合成相关基因及其调控机制进行了广泛的探索,遗传操作和酶学表征,这在这篇综述中进行了总结。
    Acarbose is widely used as α-glucosidase inhibitor in the treatment of type Ⅱ diabetes. Actinoplanes sp. is used for industrial production of acarbose. As a secondary metabolite, the biosynthesis of acarbose is quite complex. In addition to acarbose, a few acarbose structural analogs are also accumulated in the culture broth of Actinoplanes sp., which are hard to remove. Due to lack of systemic understanding of the biosynthesis and regulation mechanisms of acarbose and its structural analogs, it is difficult to eliminate or reduce the biosynthesis of the structural analogs. Recently, the advances in omics technologies and molecular biology have facilitated the investigations of biosynthesis and regulatory mechanisms of acarbose and its structural analogs in Actinoplanes sp.. The genes involved in the biosynthesis of acarbose and its structural analogs and their regulatory mechanism have been extensively explored by using bioinformatics analysis, genetic manipulation and enzymatic characterization, which is summarized in this review.
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  • 文章类型: Journal Article
    This paper briefly reviews the role of hypermethylation of host cell genes in cervical carcinogenesis and discusses potential clinical applications of methylation analysis in the management of high-risk HPV (hrHPV) -positive women. We argue that methylation assays can be used: 1. for primary triage of hrHPV-positive women to detect cervical cancer and advanced cervical intraepithelial neoplasia (CIN); 2. as secondary triage for women with minor cytological abnormalities to identify those with the highest risk of CIN3 or worse; 3. as exit test for women leaving the screening programme to identify cervical cancer and advanced CIN; and 4. to support management of CIN. TWEETABLE ABSTRACT: This paper discusses potential clinical applications of DNA methylation analysis in the management of women with a high-risk HPV infection.
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  • 文章类型: Journal Article
    菊花(ChryranthemummorifoliumRamat。)是世界范围内具有应用价值的领先花卉。开发具有新花色和形状等新颖特征的菊花新品种,工厂架构,开花时间,采后质量,以时间和成本有效的方式耐受生物和非生物胁迫是育种者的最终目标。已采用各种育种策略来改善上述性状,从传统技术,包括杂交和突变育种,一系列的分子育种方法,包括转基因技术,基因组编辑,和标记辅助选择(MAS)。此外,高通量技术最近的广泛进步,尤其是基因组学,转录组学,蛋白质组学,代谢组学,和微生物,它们统称为组学平台,导致了大量数据的收集。这些组学数据与表型信息的整合将能够鉴定负责重要性状的基因/途径。为了加速菊花的育种,已经进行了一些尝试来使用新兴的分子和组学方法。然而,将这些研究的结果应用于实际的菊花育种仍然是一个相当大的挑战,主要是由于该物种的高杂合性和多倍体。本文总结了传统和现代分子育种方法以及新兴组学技术的最新成就,并讨论了它们在改善菊花农艺和园艺特性方面的未来应用。
    Chrysanthemum (Chrysanthemum morifolium Ramat.) is a leading flower with applied value worldwide. Developing new chrysanthemum cultivars with novel characteristics such as new flower colors and shapes, plant architectures, flowering times, postharvest quality, and biotic and abiotic stress tolerance in a time- and cost-efficient manner is the ultimate goal for breeders. Various breeding strategies have been employed to improve the aforementioned traits, ranging from conventional techniques, including crossbreeding and mutation breeding, to a series of molecular breeding methods, including transgenic technology, genome editing, and marker-assisted selection (MAS). In addition, the recent extensive advances in high-throughput technologies, especially genomics, transcriptomics, proteomics, metabolomics, and microbiomics, which are collectively referred to as omics platforms, have led to the collection of substantial amounts of data. Integration of these omics data with phenotypic information will enable the identification of genes/pathways responsible for important traits. Several attempts have been made to use emerging molecular and omics methods with the aim of accelerating the breeding of chrysanthemum. However, applying the findings of such studies to practical chrysanthemum breeding remains a considerable challenge, primarily due to the high heterozygosity and polyploidy of the species. This review summarizes the recent achievements in conventional and modern molecular breeding methods and emerging omics technologies and discusses their future applications for improving the agronomic and horticultural characteristics of chrysanthemum.
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  • 文章类型: Journal Article
    在模型生物和人类以及几种牲畜物种中,已经记录了对特定分类群丰度的遗传控制以及它们与能量平衡和生长的联系。目前正在猪中研究相同机制的初步证据。未来的研究应该扩大这些结果,并引出猪肠道微生物组群体的遗传控制程度及其与生长效率的关系。在宿主与其宏基因组之间的界面上寻求更有效的猪取决于中心假设,即肠道微生物组是所有生物体生长变异性的重要组成部分。猪不逃避这个一般规则,确定生长过程中宿主与其肠道微生物群之间相互作用的重要性可能会改变游戏规则,从而实现可持续和有效的瘦肉生产。标准取样方案,测序技术,生物信息学管道和分析方法对于跨实验和群体的结果的可移植性至关重要。同样,如果要常规利用微生物组作为选择的辅助手段,则表征和解释时间和空间变异性将是必要的步骤。
    The existence of genetic control over the abundance of particular taxa and the link of these to energy balance and growth has been documented in model organisms and humans as well as several livestock species. Preliminary evidence of the same mechanisms is currently under investigation in pigs. Future research should expand these results and elicit the extent of genetic control of the gut microbiome population in swine and its relationship with growth efficiency. The quest for a more efficient pig at the interface between the host and its metagenome rests on the central hypothesis that the gut microbiome is an essential component of the variability of growth in all living organisms. Swine do not escape this general rule, and the identification of the significance of the interaction between host and its gut microbiota in the growth process could be a game-changer in the achievement of sustainable and efficient lean meat production. Standard sampling protocols, sequencing techniques, bioinformatic pipelines and methods of analysis will be paramount for the portability of results across experiments and populations. Likewise, characterizing and accounting for temporal and spatial variability will be a necessary step if microbiome is to be utilized routinely as an aid to selection.
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  • 文章类型: Journal Article
    Resistance to colistin, mediated by chromosomal mutations and more recently, by plasmid-borne mcr genes, is increasingly being reported in bacterial isolates taken from humans, animals, farms, foods, and the environment. To easily identify and contain this quickly spreading menace, efficient diagnostics that are cheaper, faster, simpler, sensitive, and specific have become indispensable and urgently necessary. A thorough and systematic review of the literature available at Pubmed, ScienceDirect and Web of Science was thus undertaken to identify articles describing novel and efficient colistin resistance- and mcr gene-detecting methods. From the final 23 studies included in this review, both phenotypic and molecular tests were found. The phenotypic tests consisted of novel culture media viz., SuperPolymyxin™, CHROMagar COL-APSE and LBJMR media, commercial automated MIC-determining instruments such as MICRONAUT-S, Vitek 2, BD Phoenix, Sensititre and MicroScan, and novel assays such as Colistin MAC test, Colispot, rapid polymxin NP test (RPNP), alteration of Zeta potential, modified RPNP test, MICRONAUT-MIC Strip, MIC Test Strip, UMIC System, and Sensitest™ Colistin. Molecular diagnostics consisted of the CT103XL microarray, eazyplex® SuperBug kit, and Taqman® /SYBR Green® real-time PCR assays, with 100% sensitivity and specificity plus a shorter turnaround time (<3 hr). Based on the sensitivity, specificity, cost, required skill and turnaround time, the RPNP test and/or novel culture media is recommended for under-resourced laboratories while the Multiplex PCR or Taqman® /SYBR Green® real-time PCR assay alongside the RPNP or novel culture media is suggested for well-resourced ones.
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  • 文章类型: Journal Article
    细菌和古细菌的适应性免疫系统,成簇的定期间隔短回文重复,CRISPR-associate(CRISPR/Cas),作为一种强大的基因编辑工具,由于其易用性和成本效益,在基因组研究领域得到了广泛的应用。CRISPR/Cas的性能依赖于精心设计的单向导RNA(sgRNA),因此,已经开发了许多生物信息学工具来帮助设计高活性和特异性的sgRNA。这些工具的设计规格各不相同,参数,基因组等等。为了帮助研究人员选择合适的工具,我们回顾了各种sgRNA设计工具,重点研究了其在目标效率预测模型和脱靶检测算法。
    The adaptive immunity system in bacteria and archaea, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-associate (CRISPR/Cas), has been adapted as a powerful gene editing tool and got a broad application in genome research field due to its ease of use and cost-effectiveness. The performance of CRISPR/Cas relies on well-designed single-guide RNA (sgRNA), so a lot of bioinformatic tools have been developed to assist the design of highly active and specific sgRNA. These tools vary in design specifications, parameters, genomes and so on. To help researchers to choose their proper tools, we reviewed various sgRNA design tools, mainly focusing on their on-target efficiency prediction model and off-target detection algorithm.
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  • 文章类型: Journal Article
    Giardia duodenalis, Cryptosporidium spp. and Toxoplasma gondii are protozoan parasites that have been highlighted as emerging foodborne pathogens by the Food and Agriculture Organization of the United Nations and the World Health Organization. According to the European Food Safety Authority, 4786 foodborne and waterborne outbreaks were reported in Europe in 2016, of which 0.4% were attributed to parasites including Cryptosporidium, Giardia and Trichinella. Until 2016, no standardized methods were available to detect Giardia, Cryptosporidium and Toxoplasma (oo)cysts in food. Therefore, no regulation exists regarding these biohazards. Nevertheless, considering their low infective dose, ingestion of foodstuffs contaminated by low quantities of these three parasites can lead to human infection. To evaluate the risk of protozoan parasites in food, efforts must be made towards exposure assessment to estimate the contamination along the food chain, from raw products to consumers. This requires determining: (i) the occurrence of infective protozoan (oo)cysts in foods, and (ii) the efficacy of control measures to eliminate this contamination. In order to conduct such assessments, methods for identification of viable (i.e. live) and infective parasites are required. This review describes the methods currently available to evaluate infectivity and viability of G. duodenalis cysts, Cryptosporidium spp. and T. gondii oocysts, and their potential for application in exposure assessment to determine the presence of the infective protozoa and/or to characterize the efficacy of control measures. Advantages and limits of each method are highlighted and an analytical strategy is proposed to assess exposure to these protozoa.
    UNASSIGNED: Estimation de la viabilité et infectiosité des stades (kystes et oocystes) de Giardia duodenalis, Cryptosporidium spp. et Toxoplasma gondii transmis par la nourriture et l’eau : une revue des méthodes.
    UNASSIGNED: Giardia duodenalis, Cryptosporidium spp. et Toxoplasma gondii sont des parasites protozoaires qui ont été soulignés comme agents pathogènes émergents dans les aliments par l’Organisation des Nations Unies pour l’alimentation et l’agriculture et l’Organisation Mondiale de la Santé. Selon l’Autorité Européenne de Sécurité des Aliments, 4786 épidémies d’origine alimentaire et hydrique ont été enregistrées en Europe en 2016, dont 0.4% ont été attribuées à des parasites, incluant Cryptosporidium, Giardia et Trichinella. Jusqu’en 2016, aucune méthode standardisée n’était disponible pour détecter les kystes de Giardia et les oocystes de Cryptosporidium et Toxoplasma dans les aliments. Aucune réglementation n’est donc proposée concernant ces dangers. Cependant, compte tenu de leur faible dose infectieuse, l’ingestion d’une quantité d’aliments faiblement contaminés peut entraîner une infection de l’homme. Pour évaluer le risque lié aux protozoaires dans les aliments, des efforts doivent être faits dans l’évaluation de l’exposition pour estimer la contamination le long de la chaîne alimentaire, depuis la matière première jusqu’aux consommateurs. Cette évaluation nécessite de déterminer : (i) la prévalence de parasites infectieux dans les aliments, (ii) l’efficacité des mesures de maîtrise pour éliminer cette contamination. Pour mener une telle évaluation, des méthodes capables d’identifier des parasites viables (vivants) et infectieux sont requises. Cette revue décrit les méthodes actuellement disponibles permettant d’évaluer l’infectiosité et la viabilité des kystes de G. duodenalis et des oocystes de Cryptosporidium spp. et T. gondii, et leur potentiel pour être appliquées dans l’évaluation de l’exposition pour déterminer la présence de parasites infectieux et/ou caractériser l’efficacité des mesures de maîtrise. Les avantages et limites de chaque méthode sont présentés et une stratégie d’analyses est proposée pour évaluer l’exposition à ces protozoaires.
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  • 文章类型: Journal Article
    The Feulgen reaction has been proposed by Robert Feulgen and Heinrich Rossenbeck for the identification of DNA nearly a hundred years ago. Since then, many other applications of this cytochemical/topochemical procedure at qualitative and quantitative level have been proposed in relation to DNA and its role in chromatin in human, animal and plant cells. In this article, we briefly review some fundamental aspects of the Feulgen reaction and current applications of such a method in studies of altered chromatin texture, including its association with or preceding changes in transcriptional activities and effect on epigenetic marks. Further perspectives on the use of the Feulgen reaction will depend of the proposal of innovative biological questions in which its reveals appropriate.
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