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1,183 results for “Insertion”
Data from: Transcriptionally active LTR retrotransposons in Eucalyptus genus are differentially expressed and insertionally polymorphic
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African cichlid fish genera (Pseudocrenilabrinae): counts of vertebrae, supraneurals, dorsal and anal pterygiophores; pterygiophore insertion patterns
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Data from: Forward genetic screening for regulators involved in cholesterol synthesis using validation-based insertional mutagenesis
Somatic cell genetics is a powerful approach for unraveling the regulatory mechanism of cholesterol metabolism. However, it is difficult to identify the mutant gene(s) due to cells are usually mutagenized chemically or physically. To identify important genes controlling cholesterol biosynthesis, an unbiased forward genetics approach named validation-based insertional mutagenesis (VBIM) system was used to isolate and characterize the 25-hydroxycholesterol (25-HC)-resistant and SR-12813-resisitant mutants. Here we report that five mutant cell lines were isolated. Among which, four sterol-resistant mutants either contain a truncated NH2-terminal domain of sterol regulatory element-binding protein (SREBP)-2 terminating at amino acids (aa) 400, or harbor an overexpressed SREBP cleavage-activating protein (SCAP). Besides, one SR-12813 resistant mutant was identified to contain a truncated COOH-terminal catalytic domain of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase). This study demonstrates that the VBIM system can be a powerful tool to screen novel regulatory genes in cholesterol biosynthesis.
Data from: Evidence for amino acid snorkeling from a high-resolution, in vivo analysis of Fis1 tail anchor insertion at the mitochondrial outer membrane
Proteins localized to mitochondria by a carboxyl-terminal tail anchor (TA) play roles in apoptosis, mitochondrial dynamics, and mitochondrial protein import. To reveal characteristics of TAs that may be important for mitochondrial targeting, we focused our attention upon the TA of the Saccharomyces cerevisiae Fis1 protein. Specifically, we generated a library of Fis1p TA variants fused to the Gal4 transcription factor, then, using next-generation sequencing, revealed which Fis1p TA mutations inhibited membrane insertion and allowed Gal4p activity in the nucleus. Prompted by our global analysis, we subsequently analyzed the ability of individual Fis1p TA mutants to localize to mitochondria. Our findings suggest that the membrane-associated domain of the Fis1p TA may be bipartite in nature, and we encountered evidence that the positively charged patch at the carboxyl-terminus of Fis1p is required for both membrane insertion and organelle specificity. Furthermore, lengthening or shortening of the Fis1p TA by up to three amino acids did not inhibit mitochondrial targeting, arguing against a model in which TA length directs insertion of TAs to distinct organelles. Most importantly, positively charged residues were more acceptable at several positions within the membrane-associated domain of the Fis1p TA than negatively charged residues. These findings, emerging from the first high-resolution analysis of an organelle targeting sequence by deep mutational scanning, provide strong, in vivo evidence that lysine and arginine can "snorkel," or become stably incorporated within a lipid bilayer by placing terminal charges of their side chains at the membrane interface.
Data from: Mining for single nucleotide polymorphisms and insertions / deletions in expressed sequence tag libraries of oil palm
The oil palm is a tropical oil bearing tree. Recently EST-derived SNPs and SSRs are a free by-product of the currently expanding EST (Expressed Sequence Tag) data bases. The development of high-throughput methods for the detection of SNPs (Single Nucleotide Polymorphism) and small indels (insertion / deletion) has led to a revolution in their use as molecular markers. Available (5452) Oil palm EST sequences were mined from dbEST of NCBI. CAP3 program was used to assemble EST sequences into contigs. Candidate SNPs and Indel polymorphisms were detected using the perl script auto_snip version 1.0 which has used 576 ESTs for detecting SNPs and Indel sites. We found 1180 SNP sites and 137 indel polymorphisms with frequency 1.36 SNPs / 100 bp. Among the six tissues from which the EST libraries had been generated, mesocarp had high frequency of 2.91 SNPs and indels per 100 bp whereas the zygotic embryos had lowest frequency of 0.15 per 100 bp. We also used the Shannon index to analyze the proportion of ten possible types of SNP/indels. ESTs from tissues of normal apex showed highest values of Shannon index (0.60) whereas abnormal apex had least value (0.02). The present report deals the use of Shannon index for comparing SNP/ indel frequencies mined from ESTlibraries and also confirm that the frequency of SNP occurrence in oil palm to use them as markers for genetic studies.
Data from: Plastome sequencing of ten nonmodel crop species uncovers a large insertion of mitochondrial DNA in cashew
In plant evolution, intracellular gene transfer (IGT) is a prevalent, ongoing process. While nuclear and mitochondrial genomes are known to integrate foreign DNA via IGT and horizontal gene transfer (HGT), plastid genomes (plastomes) have resisted foreign DNA incorporation and only recently has IGT been uncovered in the plastomes of a few land plants. In this study, we completed plastome sequences for l0 crop species and describe a number of structural features including variation in gene and intron content, inversions, and expansion and contraction of the inverted repeat (IR). We identified a putative rpl22 in cinnamon (Cinnamomum verum J. Presl) and other sequenced Lauraceae and an apparent functional transfer of rpl23 to the nucleus of quinoa (Chenopodium quinoa Willd.). In the orchard tree cashew (Anacardium occidentale L.), we report the insertion of an ∼6.7-kb fragment of mitochondrial DNA into the plastome IR. BLASTn analyses returned high identity hits to mitogenome sequences including an intact ccmB open reading frame. Using three plastome markers for five species of Anacardium, we generated a phylogeny to investigate the distribution and timing of the insertion. Four species share the insertion, suggesting that this event occurred <20 million yr ago in a single clade in the genus. Our study extends the observation of mitochondrial to plastome IGT to include long-lived tree species. While previous studies have suggested possible mechanisms facilitating IGT to the plastome, more examples of this phenomenon, along with more complete mitogenome sequences, will be required before a common, or variable, mechanism can be elucidated.
An amphipathic helical insert drives interaction of Fibrillins with plastoglobule lipid droplets
<p>Structures and trajectory files for molecular dynamics protein-membrane binding simulations</p>
COMPLEX SENTENCE WITH PARENTHESIS INSERT COMPONENT IN THE LANGUAGE AND ITS LINGUOCULTUROLOGICAL FEATURES
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Long-chain lipids facilitate insertion of large nanoparticles into membranes of small unilamellar vesicles
<div> <p>DLS data and Cryo Images of SUVs-QDs</p> <p> </p> </div>
Insertion sequences and other mobile elements associated with antibiotic resistance genes in Enterococcus isolates from an inpatient with prolonged bacteremia.
<p>Insertion sequences (ISs) and other transposable elements are associated with the mobilization of antibiotic resistance determinants and the modulation of pathogenic characteristics. In this work, we aimed to investigate the association between ISs and antibiotic resistance genes, and their role in dissemination and modification of the antibiotic resistant phenotype. To that end, we leveraged fully resolved <em>Enterococcus faecium</em> and <em>Enterococcus faecalis</em> genomes of isolates collected over five days from an inpatient with prolonged bacteremia. Isolates from both species harbored similar IS family content but showed significant species-dependent differences in copy number and arrangements of ISs throughout their replicons. Here, we describe two inter-specific IS-mediated recombination events and IS-mediated excision events in plasmids of <em>E. faecium</em> isolates. We also characterize a novel arrangement of the ISs in a Tn1546-like transposon in <em>E. faecalis</em> isolates likely implicated in a vancomycin genotype-phenotype discrepancy. Furthermore, an extended analysis revealed a novel association between daptomycin resistance mutations in <em>liaSR</em> genes and a putative composite transposon in<em> E. faecium</em>, offering a new paradigm for the study of daptomycin resistance and novel insights into the dissemination of daptomycin resistance. In conclusion, our study highlights the role ISs and other transposable elements play in the rapid adaptation and response to clinically relevant stresses such as aggressive antibiotic treatment in enterococci.</p>
Text-fig. 4. SRXTM images of Miranthus elegans gen. et sp. nov.; Mira locality, Portugal. a, b: Transverse (a) and longitudinal (b) sections of flower showing the ovary wall (ow) and numerous ovules (ov) borne on the mushroom-shaped central globose placenta (pl, blue); placenta shown as a voltex rendering added to orthoslices; note the large openings in the floral tissue (asterisks) interpreted as schizogenous secretory cavities. c, d: Longitudinal (c) and transverse (d) sections of flower (c, volume rendering cut between orthoslice yz0970-1005; d, volume rendering cut between orthoslice xy0780-0820) showing semiinferior ovary (ow, ovary wall) with sepals inserted at the rim of the hypanthium, central column (cc) with mushroom-shaped globose placenta (pl) bearing numerous ovules (ov). Specimens, Mira 100-S153146 (a, b), Mira 100-S170155 (c, d, holotype). Scale bars = 600 µm (a–d). in Early Flowers Of Primuloid Ericales From The Late Cretaceous Of Portugal And Their Ecological And Phytogeographic Implications
Text-fig. 4. SRXTM images of Miranthus elegans gen. et sp. nov.; Mira locality, Portugal. a, b: Transverse (a) and longitudinal (b) sections of flower showing the ovary wall (ow) and numerous ovules (ov) borne on the mushroom-shaped central globose placenta (pl, blue); placenta shown as a voltex rendering added to orthoslices; note the large openings in the floral tissue (asterisks) interpreted as schizogenous secretory cavities. c, d: Longitudinal (c) and transverse (d) sections of flower (c, volume rendering cut between orthoslice yz0970-1005; d, volume rendering cut between orthoslice xy0780-0820) showing semiinferior ovary (ow, ovary wall) with sepals inserted at the rim of the hypanthium, central column (cc) with mushroom-shaped globose placenta (pl) bearing numerous ovules (ov). Specimens, Mira 100-S153146 (a, b), Mira 100-S170155 (c, d, holotype). Scale bars = 600 µm (a–d).
Text-fig. 2. Caryanthus sp., no. F03286, locality Zliv-Řídká Blana A – Inflorescence, three bisexual flowers / fruits, central flower inserted slightly higher than the two lateral ones, scale bar 0.5 mm; B – Lateral flowers arranged at an angle of about 45˚ relative to each other, hp. – hypanthium with grooves, scale bar 0.5 mm; C – Epidermal cells of hypanthium, scale bar 0.1 mm; D – Detail of epidermal cells, scale bar 50 µm; E – Lateral flower, basal parts of six filaments and basal parts of two anthers styles, f – filaments, s – styles, scale bar 0.3 mm; F – Central flower, scale bar 0.3 mm; G – Stoma, scale bar 30 µm. in A New Record Of The Genus Caryanthus From The Cretaceous Of South Bohemia (Czech Republic)
Text-fig. 2. Caryanthus sp., no. F03286, locality Zliv-Řídká Blana A – Inflorescence, three bisexual flowers / fruits, central flower inserted slightly higher than the two lateral ones, scale bar 0.5 mm; B – Lateral flowers arranged at an angle of about 45˚ relative to each other, hp. – hypanthium with grooves, scale bar 0.5 mm; C – Epidermal cells of hypanthium, scale bar 0.1 mm; D – Detail of epidermal cells, scale bar 50 µm; E – Lateral flower, basal parts of six filaments and basal parts of two anthers styles, f – filaments, s – styles, scale bar 0.3 mm; F – Central flower, scale bar 0.3 mm; G – Stoma, scale bar 30 µm.
Raw data of I-V curves for Co 2ML insertion in P state.zip
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Data from: High-throughput genotyping of green algal mutants reveals random distribution of mutagenic insertion sites and endonucleolytic cleavage of transforming DNA
A high-throughput genetic screening platform in a single-celled photosynthetic eukaryote would be a transformative addition to the plant biology toolbox. Here, we present ChlaMmeSeq (Chlamydomonas MmeI-based insertion site Sequencing), a tool for simultaneous mapping of tens of thousands of mutagenic insertion sites in the eukaryotic unicellular green alga Chlamydomonas reinhardtii. We first validated ChlaMmeSeq by in-depth characterization of individual insertion sites. We then applied ChlaMmeSeq to a mutant pool and mapped 11,478 insertions, covering 39% of annotated protein coding genes. We observe that insertions are distributed in a manner largely indistinguishable from random, indicating that mutants in nearly all genes can be obtained efficiently. The data reveal that sequence-specific endonucleolytic activities cleave the transforming DNA and allow us to propose a simple model to explain the origin of the poorly understood exogenous sequences that sometimes surround insertion sites. ChlaMmeSeq is quantitatively reproducible, enabling its use for pooled enrichment screens and for the generation of indexed mutant libraries. Additionally, ChlaMmeSeq allows genotyping of hits from Chlamydomonas screens on an unprecedented scale, opening the door to comprehensive identification of genes with roles in photosynthesis, algal lipid metabolism, the algal carbon-concentrating mechanism, phototaxis, the biogenesis and function of cilia, and other processes for which C. reinhardtii is a leading model system.
Data from: Entire plastid phylogeny of the carrot genus (Daucus, Apiaceae): Concordance with nuclear data and mitochondrial and nuclear DNA insertions to the plastid
PREMISE OF THE STUDY: We explored the phylogenetic utility of entire plastid DNA sequences in Daucus and compared the results with prior phylogenetic results using plastid and nuclear DNA sequences. METHODS: We used Illumina sequencing to obtain full plastid sequences of 37 accessions of 20 Daucus taxa and outgroups, analyzed the data with phylogenetic methods, and examined evidence for mitochondrial DNA transfer to the plastid (DcMP). KEY RESULTS: Our phylogenetic trees of the entire data set were highly resolved, with 100% bootstrap support for most of the external and many of the internal clades, except for the clade of D. carota and its most closely related species D. syrticus. Subsets of the data, including regions traditionally used as phylogenetically informative regions, provide various degrees of soft congruence with the entire data set. There are areas of hard incongruence, however, with phylogenies using nuclear data. We extended knowledge of a mitochondrial to plastid DNA insertion sequence previously named DcMP and identified the first instance in flowering plants of a sequence of potential nuclear genome origin inserted into the plastid genome. There is a relationship of inverted repeat junction classes and repeat DNA to phylogeny, but no such relationship with nonsynonymous mutations. CONCLUSIONS: Our data have allowed us to (1) produce a well-resolved plastid phylogeny of Daucus, (2) evaluate subsets of the entire plastid data for phylogeny, (3) examine evidence for plastid and nuclear DNA phylogenetic incongruence, and (4) examine mitochondrial and nuclear DNA insertion into the plastid.
GraffiTE: a Unified Framework to Analyze Transposable Element Insertion Polymorphisms using Genome-graphs
<p>This archive contains the larger datasets associated with the GraffiTE project. </p> <p>See: https://github.com/cgroza/GraffiTE</p>
Evaluation of Resin Composite Posterior Restorations Using Modeling Resin Insertion Technique in Class I Lesions
ClinicalTrials.gov study NCT05801458. IPD Sharing: Not stated. Countries: 0. Publications: 19.
Assessing Dextenza Insert After SMILE Procedure
ClinicalTrials.gov study NCT04380857. IPD Sharing: NO. Countries: 1. Publications: 0.
Long Term Follow-up of the VisAbility™ Micro Insert System for Presbyopic Patients
ClinicalTrials.gov study NCT03811249. IPD Sharing: NO. Countries: 1. Publications: 0.
An Open-label Extension Study to Evaluate the Safety of the 13 mg Bimatoprost Ocular Insert
ClinicalTrials.gov study NCT02537015. IPD Sharing: Not stated. Countries: 1. Publications: 0.
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