DNA Segments Research Articles

Description of Cryptaphelenchus recticaudatus n. sp. (Aphelenchoidea: Ektaphelenchinae) in Pinus elliottii from the USA

Summary Cryptaphelenchus recticaudatus n. sp. is described and illustrated in Pinus elliottii from the USA. The new species is characterised by the female body 310-431 μm long with distinctly annulated cuticle, lateral fields with four lines, lip region separated from the body by a shallow depression, delicate stylet with small knobs, post-vulval uterine sac short, and rectum and anus invisible. Males are 228-314 μm long, spicules 9.8-12.4 μm long with a well-developed and broad condylus, and seven caudal papillae arranged as a single (P1) and pair (P2) of precloacal papillae plus two pairs of postcloacal papillae. Based upon the general female morphology, the new species most closely resembles C. baujardi and C. iranicus. The morphological differences with the aforementioned species and other species of the genus are discussed. The phylogenetic analyses based on small (SSU) and large subunit (LSU) D2-D3 expansion segments of ribosomal DNA of different individuals of the new species revealed that the new species fell into the Cryptaphelenchus clade in both SSU and LSU trees. The monophyly of the genus was retained after adding newly generated sequences of the new species.

MAJOR VIRULENCE FACTORS AND PATHOGENICITY ISLANDS IN PATHOGENIC CLOSTRIDIUM SPECIES

Clostridia are obligately anaerobic, spore-forming bacilli that, at least in the early stages of growth, stain gramme positive. Clostridia produce greater toxins than any other bacterium genus, and pathogenic clostridia are typically diagnosed by their unique toxins. Clostridium spp. has been found to have more than 20 toxins and other extracellular proteins that contribute to virulence, such as spread factors and proteolytic enzymes. Botulinum and tetanus toxins are the most potent poisons ever discovered. Clostridium botulinum neurotoxins are the most potent acute toxins identified, and they are the cause of the neuroparalytic illness botulism. Such toxins work by inhibiting presynaptic nerve terminal neurotransmission in the peripheral and central nervous systems. Other clostridia toxins have different modes of action, such as tissue destruction, hemolysis, diarrhoea, or generating an overactive immunological response in the recipient. On non-integrative lysogenic bacteriophages or plasmids, the genes coding for numerous clostridial toxins are found. Protein secretory processes in Clostridia are poorly understood. It has remained a mystery as to how the tetanus toxin, which lacks a normal N-terminal signal peptide, is exported until today. Typical PAI are DNA segments that are found in pathogenic bacteria's genomes but not in nonpathogenic strains of the same or similar species.

Detecting inversions in routine molecular diagnosis in MMR genes.

Inversions, i.e. a change in orientation of a segment of DNA, are a recognized cause of human diseases which remain overlooked due to their balanced nature. Inversions can have severe or more subtle impacts on gene expression. We describe two families that exemplify these aspects and underline the need for inversion detection in routine diagnosis. The first family (F1) displayed a sibship with two constitutional mismatch repair deficiency patients and a family history of colon cancer in the paternal branch. The second family (F2) displayed a severe history of Lynch syndrome. These families were analyzed using a whole gene panel (WGP) strategy i.e. including colon cancer genes with their intronic and flanking genomic regions. In F1, a PMS2 inversion encompassing the promoter region to intron 1 and a PMS2 splice variant were found in the maternal and paternal branch, respectively. In F2, we described the first MSH6 inversion, involving the 5' part of MSH6 and the 3' part of the nearby gene ANXA4. Inversion detection mandates genomic sequencing, but makes a valuable contribution to the diagnostic rate. WGP is an attractive strategy as it maximizes the detection power on validated genes and keeps sufficient depth to detect de novo events.

Propagation of Recombinant Genes through Complex Microbiomes with Synthetic Mini-RP4 Plasmid Vectors.

The promiscuous conjugation machinery of the Gram-negative plasmid RP4 has been reassembled in a minimized, highly transmissible vector for propagating genetically encoded traits through diverse types of naturally occurring microbial communities. To this end, the whole of the RP4-encoded transfer determinants (tra, mob genes, and origin of transfer oriT) was excised from their natural context, minimized, and recreated in the form of a streamlined DNA segment borne by an autoselective replicon. The resulting constructs (the pMATING series) could be self-transferred through a variety of prokaryotic and eukaryotic recipients employing such a rationally designed conjugal delivery device. Insertion of GFP reporter into pMATING exposed the value of this genetic tool for delivering heterologous genes to both specific mating partners and complex consortia (e.g., plant/soil rhizosphere). The results accredited the effective and functional transfer of the recombinant plasmids to a diversity of hosts. Yet the inspection of factors that limit interspecies DNA transfer in such scenarios uncovered type VI secretion systems as one of the factual barriers that check otherwise high conjugal frequencies of tested RP4 derivatives. We argue that the hereby presented programming of hyperpromiscuous gene transfer can become a phenomenal asset for the propagation of beneficial traits through various scales of the environmental microbiome.

Anion-specific structure and stability of guanidinium-bound DNA origami

While the folding of DNA into rationally designed DNA origami nanostructures has been studied extensively with the aim of increasing structural diversity and introducing functionality, the fundamental physical and chemical properties of these nanostructures remain largely elusive. Here, we investigate the correlation between atomistic, molecular, nanoscopic, and thermodynamic properties of DNA origami triangles. Using guanidinium (Gdm) as a DNA-stabilizing but potentially also denaturing cation, we explore the dependence of DNA origami stability on the identity of the accompanying anions. The statistical analyses of atomic force microscopy (AFM) images and circular dichroism (CD) spectra reveals that sulfate and chloride exert stabilizing and destabilizing effects, respectively, already below the global melting temperature of the DNA origami triangles. We identify structural transitions during thermal denaturation and show that heat capacity changes ΔCp determine the temperature sensitivity of structural damage. The different hydration shells of the anions and their potential to form Gdm+ ion pairs in concentrated salt solutions modulate ΔCp by altered wetting properties of hydrophobic DNA surface regions as shown by molecular dynamics simulations. The underlying structural changes on the molecular scale become amplified by the large number of structurally coupled DNA segments and thereby find nanoscopic correlations in AFM images.

Polymer architecture orchestrates the segregation and spatial organization of replicating E. coli chromosomes in slow growth.

The mechanism of chromosome segregation and organization in the bacterial cell cycle of E. coli is one of the least understood aspects in its life cycle. The E. coli chromosome is often modelled as a bead spring ring polymer. We introduce cross-links in the DNA-ring polymer, resulting in the formation of loops within each replicating bacterial chromosome. We use simulations to show that the chosen polymer-topology ensures its self-organization along the cell long-axis, such that various chromosomal loci get spatially localized as seen in vivo. The localization of loci arises due to entropic repulsion between polymer loops within each daughter DNA confined in a cylinder. The cellular addresses of the loci in our model are in fair agreement with those seen in experiments as given in J. A. Cass et al., Biophys. J., 2016, 110, 2597-2609. We also show that the adoption of such modified polymer architectures by the daughter DNAs leads to an enhanced propensity of their spatial segregation. Secondly, we match other experimentally reported results, including observation of the cohesion time and the ter-transition. Additionally, the contact map generated from our simulations reproduces the macro-domain like organization as seen in the experimentally obtained Hi-C map. Lastly, we have also proposed a plausible reconciliation of the 'Train Track' and the 'Replication Factory' models which provide conflicting descriptions of the spatial organization of the replication forks. Thus, we reconcile observations from complementary experimental techniques probing bacterial chromosome organization.

A New Tool for the Flexible Genetic Manipulation of Geobacillus kaustophilus.

Geobacillus kaustophilus , a thermophilic Gram-positive bacterium, is an attractive host for the development of high-temperature bioprocesses. However, its reluctance against genetic manipulation by standard methodologies hampers its exploitation. Here, we describe a simple methodology in which an artificial DNA segment on the chromosome of Bacillus subtilis can be transferred via pLS20-mediated conjugation resulting in subsequent integration in the genome of G. kaustophilus. Therefore, we have developed a transformation strategy to design an artificial DNA segment on the chromosome of B. subtilis and introduce it into G. kaustophilus . The artificial DNA segment can be freely designed by taking advantage of the plasticity of the B. subtilis genome and combined with the simplicity of pLS20 conjugation transfer. This transformation strategy would adapt to various Gram-positive bacteria other than G. kaustophilus . Graphical abstract.

Bacillus subtilis 168 as a unique platform enabling synthesis and dissemination of genomes.

Bacillus subtilis Marburg 168 is a unique platform for genome engineering and genome synthesis. Genome scale DNA sequences can be synthesized by repeated integration of small DNA segments in the B. subtilis genome. The small DNA segments are collectively called dominos, and should cover the target genome. The B. subtilis strains which have been designed for use in the domino method are collectively called BGM: Bacillus subtilis Genome for Manipulation. The BGM system has been used to produce various genomes in the B. subtilis genome. The synthesized genomes have been demonstrated to be stably maintained as part of the B. subtilis genome. Instability of the synthesized genomes have been observed in genomes with Guanine plus Cytosine contents much higher or lower than that of BGM. The largest synthesized genome produced using this approach to date is that from Synecchosystis PCC6803, a photosynthetic microbe with a genome size of about 3.5 Mbp. The domino method depends on transformation, using the natural competence of B. subtilis. An alternative DNA uptake system, conjugational transfer, has been studied for the past 20 years. A self-transmissible plasmid named pLS20 has been used for the transfer and delivery of large amounts of DNA between B. subtilis. The BGM system is a unique platform for handling very large amounts of DNA from synthesis to dissemination to other cells, and has broad applications in research and practice.

The Relationship Between FokI Vitamin D Receptor Polymorphisms on The Incidence Of Diabetic Retinopathy In Type 2 Diabetes Mellitus Patients At Sanglah Hospital Denpasar Bali

Introduction: Complications of T2DM result in many organs of the body including visual disturbances (diabetic retinopathy). Polymorphisms in vitamin D receptor gene FokI, increase the risk of complications of T2DM including DR. This study aims to determine the FokI polymorphism in vitamin D receptor gene in DMT2 patients at Sanglah Hospital Denpasar Bali and to determine whether there is a relationship between vitamin D receptor polymorphisms FokI on the incidence of DR in DMT2 patients at Sanglah Hospital Denpasar Bali. Methods: Design of the research was a cross-sectional analytical study. To detect the status of the FokI polymorphism, DNA segment amplification was performed using the PCR method using a primer specific for the FokI polymorphism. Furthermore, sequencing was performed to detect the base pattern at the location of the FokI SNP. Statistical analysis was performed by using SPSS 25.sav. P-value <0.05 was considered statistically significant. Results: There were 38 samples that were eligible to be analysed and successfully sequenced. The mean age of the sample was 50.68 years with the proportion of DR and non-DR of 73.7% and 26.3%, with the majority of DR patients being male (65.8%). In our multivariate analysis, we found a significant relationship between VDR FokI polymorphism with hypertension among diabetic retinopathy patient (p=0.036; adjusted OR=18; 95% CI=1.694-191.233). Conclusion: Heterozygous (C/T) VDR Fok I polymorphism was the most frequent in this finding. There is no significant relationship between FokI polymorphisms in vitamin D receptor gene and the incidence of RD in T2DM patients at Sanglah Hospital Denpasar.

An Efficient Markerless Deletion System Suitable for the Industrial Strains of Streptomyces.

The genus Streptomyces is intensively studied due to its excellent ability to produce secondary metabolites with diverse bioactivities. In particular, adequate precursors of secondary metabolites as well as sophisticated post modification systems make some high-yield industrial strains of Streptomyces the promising chassis for the heterologous production of natural products. However, lack of efficient genetic tools for the manipulation of industrial strains, especially the episomal vector independent tools suitable for large DNA fragment deletion, makes it difficult to remold the metabolic pathways and streamline the genomes in these strains. In this respect, we developed an efficient deletion system independent of the episomal vector for large DNA fragment deletion. Based on this system, four large segments of DNA, ranging in length from 10 kb to 200 kb, were knocked out successfully from three industrial Streptomyces strains without any marker left. Notably, compared to the classical deletion system used in Streptomyces, this deletion system takes about 25% less time in our cases. This work provides a very effective tool for further genetic engineering of the industrial Streptomyces.

Analysis of molecular diversity of Capsicum spp. to identify redundant accessions and duplicates within accessions from a germplasm bank

Assessment of genetic diversity and identification of redundant accessions in seed banks are important to minimize the risk of gene loss in collections and to decrease costs in the management of germplasm banks. Current study used markers at DNA level to evaluate genetic diversity and identify whether there are duplicate accessions of Capsicum spp. in the collections of an active germplasm bank. Randomly amplified polymorphic DNA markers using touchdown-polymerase chain reaction method evaluated genetic diversity and identified duplicates in ten accessions of Capsicum spp. Higher genetic diversity has been detected in C. annuum rather than in the species C. chinense and C. baccatum. DNA amplified segments revealed high genetic divergence showing that accessions of C. annuum, C. chinense and C. baccatum were separately grouped after data analysis with three software programs. Accessions of C. baccatum var. pendulum and C. annuum var. glabriusculum were the least similar. Since rates were less than 60% between accessions of C. chinense and C. annuum, they revealed low identity of amplified DNA segments among the three Capsicum species. Redundance in four accessions of C. chinense and in five accessions of C. annuum was not detected. On the other hand, duplicates within accessions C. chinense (50%), C. annuum var. glabrisculum (5%) and C. baccatum var. pendulum (75%) were reported in the Capsicum active germplasm bank.

DNA supercoiling-mediated collective behavior of co-transcribing RNA polymerases.

Multiple RNA polymerases (RNAPs) transcribing a gene have been known to exhibit collective group behavior, causing the transcription elongation rate to increase with the rate of transcription initiation. Such behavior has long been believed to be driven by a physical interaction or ‘push’ between closely spaced RNAPs. However, recent studies have posited that RNAPs separated by longer distances may cooperate by modifying the DNA segment under transcription. Here, we present a theoretical model incorporating the mechanical coupling between RNAP translocation and the DNA torsional response. Using stochastic simulations, we demonstrate DNA supercoiling-mediated long-range cooperation between co-transcribing RNAPs. We find that inhibiting transcription initiation can slow down the already recruited RNAPs, in agreement with recent experimental observations, and predict that the average transcription elongation rate varies non-monotonically with the rate of transcription initiation. We further show that while RNAPs transcribing neighboring genes oriented in tandem can cooperate, those transcribing genes in divergent or convergent orientations can act antagonistically, and that such behavior holds over a large range of intergenic separations. Our model makes testable predictions, revealing how the mechanical interplay between RNAPs and the DNA they transcribe can govern transcriptional dynamics.

Amyloid Precursor Protein A713T Mutation in Calabrian Patients with Alzheimer's Disease: A Population Genomics Approach to Estimate Inheritance from a Common Ancestor.

Mutation A713T in the amyloid precursor protein (APP) has been linked to cases of Alzheimer’s disease (AD), cerebral amyloid angiopathy (CAA) and cerebrovascular disease. Despite its rarity, it has been observed in several families from the same geographical area, in the Calabria region in Southern Italy. Genotyping of 720,000 genome-wide SNPs with the HumanOmniExpress BeadChip was performed for six patients that were representative of apparently unrelated Calabrian families, as well as a Belgian subject of Italian descent (all with the same A713T mutation and disease). Their genomic structure and genetic relationships were analyzed. Demographic reconstruction and coalescent theory were applied to estimate the time of the most recent common ancestor (tMRCA) among patients. Results show that all A713T carriers fell into the genetic variability of Southern Italy and were not more closely related to each other than to any other healthy Calabrian individual. However, five out of seven patients shared a 1.7 Mbp-long DNA segment centered on the A713T mutation, making it possible to estimate a tMRCA for its common origin in the Calabrian region dating back over 1000 years. The analysis of affected individuals with methodologies based on human population genomics thus provides informative insights in support of clinical observations and biomedical research.

Pan-Genomic Sequencing Reveals Actionable CDKN2A/2B Deletions and Kataegis in Anaplastic Thyroid Carcinoma.

Simple SummaryAnaplastic thyroid carcinoma (ATC) is a tumor with exceedingly high mortality rates, and current treatment options are very limited once the disease has spread beyond the thyroid gland. Most of what we know of ATCs from a genetic standpoint is based upon previous analyses in which limited DNA segments are sequenced, and little is known about the so-called non-coding DNA regions. To identify novel genetic mechanisms of potential therapeutic use, we sequenced the entire genome of eight ATC cases and corresponding normal tissues. We found a recurrent deletion of two neighboring genes responsible for inhibiting cell division, and these findings may be of clinical interest as there are specific drugs available for tumors with this gene aberration. We also found regional aggregations of mutations in specific clusters, a phenomenon entitled “kataegis”, which in turn could be therapeutically relevant.Anaplastic thyroid carcinoma (ATC) is a lethal malignancy characterized by poor response to conventional therapies. Whole-genome sequencing (WGS) analyses of this tumor type are limited, and we therefore interrogated eight ATCs using WGS and RNA sequencing. Five out of eight cases (63%) displayed cyclin-dependent kinase inhibitor 2A (CDKN2A) abnormalities, either copy number loss (n = 4) or truncating mutations (n = 1). All four cases with loss of the CDKN2A locus (encoding p16 and p14arf) also exhibited loss of the neighboring CDKN2B gene (encoding p15ink4b), and displayed reduced CDKN2A/2B mRNA levels. Mutations in established ATC-related genes were observed, including TP53, BRAF, ARID1A, and RB1, and overrepresentation of mutations were also noted in 13 additional cancer genes. One of the more predominant mutational signatures was intimately coupled to the activity of Apolipoprotein B mRNA-editing enzyme, the catalytic polypeptide-like (APOBEC) family of cytidine deaminases implied in kataegis, a focal hypermutation phenotype, which was observed in 4/8 (50%) cases. We corroborate the roles of CDKN2A/2B in ATC development and identify kataegis as a recurrent phenomenon. Our findings pinpoint clinically relevant alterations, which may indicate response to CDK inhibitors, and focal hypermutational phenotypes that may be coupled to improved responses using immune checkpoint inhibitors.

In Silico Investigation of the Biological Implications of Complex DNA Damage with Emphasis in Cancer Radiotherapy through a Systems Biology Approach.

Different types of DNA lesions forming in close vicinity, create clusters of damaged sites termed as “clustered/complex DNA damage” and they are considered to be a major challenge for DNA repair mechanisms resulting in significant repair delays and induction of genomic instability. Upon detection of DNA damage, the corresponding DNA damage response and repair (DDR/R) mechanisms are activated. The inability of cells to process clustered DNA lesions efficiently has a great impact on the normal function and survival of cells. If complex lesions are left unrepaired or misrepaired, they can lead to mutations and if persistent, they may lead to apoptotic cell death. In this in silico study, and through rigorous data mining, we have identified human genes that are activated upon complex DNA damage induction like in the case of ionizing radiation (IR) and beyond the standard DNA repair pathways, and are also involved in cancer pathways, by employing stringent bioinformatics and systems biology methodologies. Given that IR can cause repair resistant lesions within a short DNA segment (a few nm), thereby augmenting the hazardous and toxic effects of radiation, we also investigated the possible implication of the most biologically important of those genes in comorbid non-neoplastic diseases through network integration, as well as their potential for predicting survival in cancer patients.

Taxonomic and genetic assessment of captive White-Handed Gibbons (Hylobateslar) in Peninsular Malaysia with implications towards conservation translocation and reintroduction programmes.

Conservation translocation and reintroduction for the purpose of repopulating and reinforcing extirpated or depleted populations has been recognised as an important conservation tool, particularly for gibbon conservation in the immediate future. Feasibility assessments involving multiple factors, including taxonomic and genetic assessment of rescued and captive gibbons, are imperative prior to translocation and reintroduction programmes. In this study, we attempt to determine the subspecies and origin of captive Hylobateslar, White-handed gibbons, from Peninsular Malaysia to assist in future translocation and reintroduction programmes. A total of 12 captive and rescued H.lar samples were analysed using the control region segment of mitochondrial DNA. Sequence analyses and phylogenetic trees constructed using neighbour-joining, maximum likelihood, Bayesian inference, and network methods congruently differentiate all 12 captive individuals used in this study from other H.lar subspecies suggesting that these individuals belong to the H.larlar subspecies. In addition, two populations of H.l.lar were observed: (1) a southern population consisting of all 12 individuals from Peninsular Malaysia, and (2) a possible northern population represented by three individuals (from previous studies), which might have originated from the region between the Isthmus of Kra, Surat Thani-Krabi depression, and Kangar-Pattani. Our findings suggest that the complete control region segment can be used to determine the subspecies and origin of captive H.lar.

Assembly of Multiple Full-Size Genes or Genomic DNA Fragments on Human Artificial Chromosomes Using the Iterative Integration System.

Human artificial chromosomes (HACs) are gene delivery vectors that have been used for decades for gene functional studies. HACs have several advantages over viral‐based gene transfer systems, including stable episomal maintenance in a single copy in the cell and the ability to carry up to megabase‐sized genomic DNA segments. We have previously developed the alphoidtetO‐HAC, which has a single gene acceptor loxP site that allows insertion of an individual gene of interest using Chinese hamster ovary (CHO) hybrid cells. The HAC, along with a DNA segment of interest, can then be transferred from donor CHO cells to various recipient cells of interest via microcell‐mediated chromosome transfer (MMCT). Here, we detail a protocol for loading multiple genomic DNA segments or genes into the alphoidtetO‐HAC vector using an iterative integration system (IIS) that utilizes recombinases Cre, ΦC31, and ΦBT. This IIS‐alphoidtetO‐HAC can be used for either serially assembling genomic loci or fragments of a large gene, or for inserting multiple genes into the same artificial chromosome. The insertions are executed iteratively, whereby each round results in the insertion of a new DNA segment of interest. This is accompanied by changes of expression of marker fluorescent proteins, which simplifies screening of correct clones, and changes of selection and counterselection markers, which constitutes an error‐proofing mechanism that removes mis‐incorporated DNA segments. In addition, the IIS‐alphoidtetO‐HAC carrying the genes can be eliminated from the cells, offering the possibility to compare the phenotypes of human cells with and without functional copies of the genes of interest. The resulting HAC molecules may be used to investigate biomedically relevant pathways or the regulation of multiple genes, and to potentially engineer synthetic chromosomes with a specific set of genes of interest. The IIS‐alphoidtetO‐HAC system is expected to be beneficial in creating multiple‐gene humanized models with the purpose of understanding complex multi‐gene genetic disorders. Published 2021. This article is a U.S. Government work and is in the public domain in the USA. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Integration of the first DNA segment of interest into the IIS‐alphoidteto‐HAC Basic Protocol 2: Integration of a second DNA segment of interest into the IIS‐alphoidteto‐HAC Basic Protocol 3: Integration of a third DNA segment of interest into the IIS‐alphoidteto‐HAC Support Protocol: Fluorescence in situ hybridization analysis for the circular IIS‐alphoidtetO‐HAC

MCKAT: a multi-dimensional copy number variant kernel association test

BackgroundCopy number variants (CNVs) are the gain or loss of DNA segments in the genome. Studies have shown that CNVs are linked to various disorders, including autism, intellectual disability, and schizophrenia. Consequently, the interest in studying a possible association of CNVs to specific disease traits is growing. However, due to the specific multi-dimensional characteristics of the CNVs, methods for testing the association between CNVs and the disease-related traits are still underdeveloped. We propose a novel multi-dimensional CNV kernel association test (MCKAT) in this paper. We aim to find significant associations between CNVs and disease-related traits using kernel-based methods.ResultsWe address the multi-dimensionality in CNV characteristics. We first design a single pair CNV kernel, which contains three sub-kernels to summarize the similarity between two CNVs considering all CNV characteristics. Then, aggregate single pair CNV kernel to the whole chromosome CNV kernel, which summarizes the similarity between CNVs in two or more chromosomes. Finally, the association between the CNVs and disease-related traits is evaluated by comparing the similarity in the trait with kernel-based similarity using a score test in a random effect model. We apply MCKAT on genome-wide CNV datasets to examine the association between CNVs and disease-related traits, which demonstrates the potential usefulness the proposed method has for the CNV association tests. We compare the performance of MCKAT with CKAT, a uni-dimensional kernel method. Based on the results, MCKAT indicates stronger evidence, smaller p-value, in detecting significant associations between CNVs and disease-related traits in both rare and common CNV datasets.ConclusionA multi-dimensional copy number variant kernel association test can detect statistically significant associated CNV regions with any disease-related trait. MCKAT can provide biologists with CNV hot spots at the cytogenetic band level that CNVs on them may have a significant association with disease-related traits. Using MCKAT, biologists can narrow their investigation from the whole genome, including many genes and CNVs, to more specific cytogenetic bands that MCKAT identifies. Furthermore, MCKAT can help biologists detect significantly associated CNVs with disease-related traits across a patient group instead of examining each subject’s CNVs case by case.

REVIEW OF DNA VACCINE APPROACHES AGAINST THE PARASITE TOXOPLASMA GONDII.

Toxoplasma gondii is an apicomplexan parasite that affects both humans and livestock. Transmitted to humans through ingestion, it is the second-leading cause of foodborne illness-related death. Currently, there exists no approved vaccine for humans or most livestock against the parasite. DNA vaccines, a type of subunit vaccine which uses segments of the pathogen's DNA to generate immunity, have shown varying degrees of experimental efficacy against infection caused by the parasite. This review compiles DNA vaccine efforts against Toxoplasma gondii, segmenting the analysis by parasite antigen, as well as a review of concomitant adjuvant usage. No single antigenic group was consistently more effective within in vivo trials relative to others.

R-Loop Tracker: Web Access-Based Tool for R-Loop Detection and Analysis in Genomic DNA Sequences

R-loops are common non-B nucleic acid structures formed by a three-stranded nucleic acid composed of an RNA–DNA hybrid and a displaced single-stranded DNA (ssDNA) loop. Because the aberrant R-loop formation leads to increased mutagenesis, hyper-recombination, rearrangements, and transcription-replication collisions, it is regarded as important in human diseases. Therefore, its prevalence and distribution in genomes are studied intensively. However, in silico tools for R-loop prediction are limited, and therefore, we have developed the R-loop tracker tool, which was implemented as a part of the DNA Analyser web server. This new tool is focused upon (1) prediction of R-loops in genomic DNA without length and sequence limitations; (2) integration of R-loop tracker results with other tools for nucleic acids analyses, including Genome Browser; (3) internal cross-evaluation of in silico results with experimental data, where available; (4) easy export and correlation analyses with other genome features and markers; and (5) enhanced visualization outputs. Our new R-loop tracker tool is freely accessible on the web pages of DNA Analyser tools, and its implementation on the web-based server allows effective analyses not only for DNA segments but also for full chromosomes and genomes.