875 resultados para 16s rRNA sequencing


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Photosynthetic eukaryotes have a critical role as the main producers in most ecosystems of the biosphere. The ongoing environmental metabarcoding revolution opens the perspective for holistic ecosystems biological studies of these organisms, in particular the unicellular microalgae that often lack distinctive morphological characters and have complex life cycles. To interpret environmental sequences, metabarcoding necessarily relies on taxonomically curated databases containing reference sequences of the targeted gene (or barcode) from identified organisms. To date, no such reference framework exists for photosynthetic eukaryotes. In this study, we built the PhytoREF database that contains 6490 plastidial 16S rDNA reference sequences that originate from a large diversity of eukaryotes representing all known major photosynthetic lineages. We compiled 3333 amplicon sequences available from public databases and 879 sequences extracted from plastidial genomes, and generated 411 novel sequences from cultured marine microalgal strains belonging to different eukaryotic lineages. A total of 1867 environmental Sanger 16S rDNA sequences were also included in the database. Stringent quality filtering and a phylogeny-based taxonomic classification were applied for each 16S rDNA sequence. The database mainly focuses on marine microalgae, but sequences from land plants (representing half of the PhytoREF sequences) and freshwater taxa were also included to broaden the applicability of PhytoREF to different aquatic and terrestrial habitats. PhytoREF, accessible via a web interface (http://phytoref.fr), is a new resource in molecular ecology to foster the discovery, assessment and monitoring of the diversity of photosynthetic eukaryotes using high-throughput sequencing.

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Photosynthetic eukaryotes have a critical role as the main producers in most ecosystems of the biosphere. The ongoing environmental metabarcoding revolution opens the perspective for holistic ecosystems biological studies of these organisms, in particular the unicellular microalgae that often lack distinctive morphological characters and have complex life cycles. To interpret environmental sequences, metabarcoding necessarily relies on taxonomically curated databases containing reference sequences of the targeted gene (or barcode) from identified organisms. To date, no such reference framework exists for photosynthetic eukaryotes. In this study, we built the PhytoREF database that contains 6490 plastidial 16S rDNA reference sequences that originate from a large diversity of eukaryotes representing all known major photosynthetic lineages. We compiled 3333 amplicon sequences available from public databases and 879 sequences extracted from plastidial genomes, and generated 411 novel sequences from cultured marine microalgal strains belonging to different eukaryotic lineages. A total of 1867 environmental Sanger 16S rDNA sequences were also included in the database. Stringent quality filtering and a phylogeny-based taxonomic classification were applied for each 16S rDNA sequence. The database mainly focuses on marine microalgae, but sequences from land plants (representing half of the PhytoREF sequences) and freshwater taxa were also included to broaden the applicability of PhytoREF to different aquatic and terrestrial habitats. PhytoREF, accessible via a web interface (http://phytoref.fr), is a new resource in molecular ecology to foster the discovery, assessment and monitoring of the diversity of photosynthetic eukaryotes using high-throughput sequencing.

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A clone showing female-specific expression was identified from an embryonic cDNA library of a mealybug, Planococcus lilacinus, In Southern blots this clone (P7) showed hybridization to genomic DNA of females, but not to that of males, However, P7 showed no hybridization to nuclei of either sex, raising the possibility that it was extrachromosomal in origin, In sectioned adult females P7 hybridized to an abdominal organ called the mycetome. The mycetome is formed by mycetocytes, which are polyploid cells originating from the polar bodies and cleavage nuclei that harbour maternally transmitted, intracellular symbionts. Electron microscopy confirmed the presence of symbionts within the mycetocytes, Sequence analysis showed that P7 is a 16S rRNA gene, confirming its prokaryotic origin, P7 transcripts are localized to one pole in young embryos but are found in the pole as well as in the germ band during later stages of development, P7 expression is detectable in young embryos of both sexes but the absence of P7 in third instar and adult males suggests that this gene, and hence the endosymbionts, are subject to sex-specific elimination. Copyright (C) 1997 Elsevier Science Ltd.

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 对大银鱼和小齿日本银鱼的线粒体细胞色素b 和16S rRNA 基因片段进行了扩增和序列测定,分析比较了2 种 间的序列差异。大银鱼2 个个体间细胞色素b 基因序列无差异,小齿日本银鱼3 个个体的序列出现了2 种单倍型;大银鱼 同小齿日本银鱼细胞色素b 序列(单倍型SB21) 之间存在86 个碱基差异(差异率21 %) ,变异较大。大银鱼、小齿日本银鱼 的16S rRNA 基因片段种内个体间无序列差异,两种间存在21 个碱基的差异(差异率5 %) ,有1 个碱基的插入/ 缺失。由 此可见,2 种银鱼间线粒体细胞色素b 基因的进化速率较快,约为16S rRNA 基因片段的4 倍。根据细胞色素b 序列数据, 推算出2 种银鱼大概在中新世晚期发生分化。

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Based on partial sequences of the 12S and 16S ribosomal RNA genes, we estimated phylogenetic relationships among brown frogs of the Rana temporaria group from China. From the phylogenetic trees obtained, we propose to include Rana zhengi in the brown frog

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A 3. 6 ns molecular dynamics simulation was carried out on the complex system of tobramycin and 16S rRNA in order to understand the speciality recognition mechanism between tobramycin and 16S rRNA at the molecular level. The results demonstrate that two l

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测定了4个种(红蹼树蛙、黑蹼树蛙、白斑小树蛙和红吸盘小树蛙)共11个种群的16S rRNA基因片段.双斑树蛙、马来棱皮树蛙、越南棱皮树蛙以及日本溪树蛙的同源序列通过GenBank检索获得.去除所有插入、缺失及模糊位点后,比对序列长度为500 bp,其中变异位点115个,简约信息位点92个.以日本溪树蛙为外群,运用Bayesian法、MP法和ML法构建了系统发育树.结果表明红蹼树蛙和白斑小树蛙在种级水平上均不是单系.红蹼树蛙海南种群与双斑树蛙亲缘关系更近,并且来自云南不同地理种群的红蹼树蛙可以分为两大支系;越南棱皮树蛙与红吸盘小树蛙聚为一支,马来棱皮树蛙嵌套在白斑小树蛙不同地理种群中.进而认为白斑小树蛙是马来棱皮树蛙的同物异名,建议将红吸盘小树蛙并入棱皮树蛙属.

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The generic allocation of Indian and Sri Lankan Philautus needs further examination. In this study, a comprehensive understanding of the phylogeny of Indian and Sri Lankan Philautus is obtained based on 125 and 16S rRNA genes. All phylogenetic analyses indicate that Indian-Sri Lankan Philautus, Philautus menglaensis, Philautus longchuanensis, and Philautus gryllus form a well supported clade, separate from Philautus of Sunda Islands that form another well supported clade representing true Philautus. This result supports the designation of the genus Pseudophilautus to accommodate the Indian and Sri Lankan species. Pseudophilautus consists of two major lineages, one comprises the majority of Indian species, Chinese species, and Southeast Asian species, and one comprises all Sri Lankan species and a few Indian species. Pseudophilautus may have originated in South Asia and dispersed into Southeast Asia and China. Based on the results, we further suggest that Philautus cf. gryllus (MNHN1997.5460) belongs to the genus Kurixalus. (C) 2010 Published by Elsevier Ltd.

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采用分子动力学方法(Molecular dynamics,MD)对托普霉素(Tobramycin)与16S rRNA的A位点复合物的特异性识别机制进行了理论模拟研究,模拟时间为3.6 ns. 结果表明,A位点中波动最大的部位是两个环外碱基A1492和A1493;tobramycin的环Ⅰ和环Ⅱ是其最保守的结构单元,可能参与了Tobramycin与16S rRNA的A位点之间的特异性识别. 另外,发现一个残存时间为3.6 ns的"结构化"水分子,它桥接了Tobramycin环Ⅱ的N3与环Ⅰ的N6′之间的氢键,稳定了Tobramycin的结构;Tobramycin周围水合密度较高的位点出现在环Ⅰ和环Ⅱ附近,这也正是晶体结构中形成较多水媒介氢键及动力学模拟中结构化水分子出现的位置. 动力学模拟证实Tobramycin与16S rRNA间的结合是大量氢键及水分子相互作用的结果,这有助于设计和开发以Tobramycin为基础,具有高亲和力及特异性的16S rRNA抑制剂.

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The sequences of the 16S rRNA genes from 38 strains of the family Thermaceae were compared by alignment analysis. The genus-specific and species-specific base substitutions or base deletions (signature positions) were found in three hypervariable regions (in the helices 6, 10 and 17). The differentiation of secondary structures of the high variable regions in the 5' end (38-497) containing several signature positions further supported the concept. Based on the comparisons of the secondary structures in the segments of 16S rRNAs, a key to the species of the family Thermaceae was proposed. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Microbiological Societies.

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To understand pharmacophore properties of pyranmycin derivatives and to design novel inhibitors of 16S rRNA A site, comparative molecular field analysis (CoMFA) approach was applied to analyze three-dimensional quantitative structure-activity relationship (3D-QSAR) of 17 compounds. AutoDock 3.0.5 program was employed to locate the orientations and conformations of the inhibitors interacting with 16S rRNA A site. The interaction mode was demonstrated in the aspects of inhibitor conformation, hydrogen bonding and electrostatic interaction. Similar binding conformations of these inhibitors and good correlations between the calculated binding free energies and experimental biological activities suggest that the binding conformations of these inhibitors derived from docking procedure were reasonable. Robust and predictive 3D-QSAR model was obtained by CoMFA with q(2) values of 0.723 and 0.993 for cross-validated and noncross-validated, respectively. The 3D-QSAR model built here will provide clear guidelines for novel inhibitors design based on the Pyranmycin derivatives against 16S rRNA A site. (c) 2005 Elsevier B.V. All rights reserved.

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分别在2004年、2005年和2006年洱海鱼腥藻水华暴发时期,分离优势种,获得藻株EH-A、EH-B和EH-C,通过形态学特征和16S rRNA基因序列分析鉴定了藻株的种类。选用藻丝的形态、气囊的存在与否、异形胞和孢子的位置、各种细胞的形状以及营养细胞、异形胞和孢子的大小等传统的分类特征描述藻株的形态。依据形态特征,初步判断这3个藻株可能为卷曲鱼腥藻(Anabaena circinalis)或A.crassa株系成员。利用16S rRNA基因序列构建邻接树分析了藻株间的系统进化关系,分析表明:藻株EH-

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对从青岚湖采集的 10属 14种河蚌的 19个样本进行了 16SrRNA的序列测定 ,并同GenBank中鄱阳湖流域相同物种河蚌的序列比较 ,分析了基于Kimura 2 parameter模型参数得到的遗传距离 ,并构建了它们的UPGMA树。结果显示 ,所有用于比较的河蚌种间遗传距离变化范围在 0 .0 2 74— 0 .2 2 90 ,平均为 0 .132 5 ,种内遗传距离在0 .0 0 34— 0 .0 0 6 8之间 ,平均为 0 .0 0 4 5 ,种间遗传距离远大于种内距离。表明以 16SrR