Nucleotide Sequence Information Research Articles

Cloning, Identification, and Functional Analysis of the Foxl2 Gene in Procambarus clarkii.

Procambarus clarkii is the most widely distributed freshwater shrimp in China, with important economic value and great potential for development. The forkheadboxL2 (Foxl2) gene has been found to be involved in the reproductive development of many crustaceans. To understand the role of the Foxl2 gene in the gonad development of P. clarkii, we designed CDS-specific primers for the P. clarkii Foxl2 (PcFoxl2) gene and cloned its CDS sequence using RT-PCR. The nucleotide and protein sequence information was then analyzed through bioinformatics analysis. The expression and subcellular localization of PcFoxl2 in various tissues were detected using qRT-PCR and in situ hybridization. The effects of PcFoxl2 knockdown on gonad development were investigated using RNA interference. The results showed that the CDS length of the PcFoxl2 gene was 1614 bp and encoded 537 amino acids. Protein sequence comparison and phylogenetic analysis showed that PcFoxl2 was the closest relative to Crayfish. qRT-PCR analysis indicated that the expression level of PcFoxl2 in the testis was significantly higher (>40 fold) than that in the ovary (p < 0.01). The in situ hybridization results showed that PcFoxl2 was expressed in both the cytoplasm and the nucleus of egg cells, and that the expression was strongest in egg cells at the early stage of yolk synthesis, while weak in the secondary oocytes. The positive signal was strongest in the spermatocyte nucleolus, while only a trace signal was observed in the cytoplasm. After interfering with the PcFoxl2 gene using dsRNA, the expression of PcFoxl2 in the RNA interference group was significantly lower than that in the control group, and this interference effect lasted for one week. Moreover, the gonad index of the experimental group was significantly lower than that of the control group (p < 0.05) after 10 days of P. clarkii cultivation following PcFoxl2 knockdown. The expression levels of the nanos and S3a genes, which are related to gonad development, decreased significantly after PcFoxl2 gene interference. The results suggest that the Foxl2 gene is involved in the growth and development of gonads, particularly in the development of testis, and is related to the early development of oocytes. This study provides a theoretical basis for the artificial breeding of P. clarkii.

Report of Phytophthora capsici (mating type A1) isolates associated with crown‐rot disease of Capsicum annuum in Guatemala

AbstractWilting and crown‐rot symptoms were observed in sweet pepper (Capsicum annuum) plants in field surveys carried out in 2021 across major vegetable‐producing areas of three municipalities of Guatemala. Phytophthora capsici (Peronosporales, Peronosporaceae) isolates were identified in association with these symptoms based upon morphological and phylogenetic analyses using nucleotide sequence information of the Cox2 mitochondrial DNA (mtDNA) region. Biological tests confirmed P. capsici mating type A1 isolates as the causal agents of the disease. Symptoms similar to those observed in the field were obtained after controlled inoculation of C. annuum ‘Tico’ seedlings. Solanum habrochaites ‘LA 404’ and C. annuum ‘Criollo de Morelos’ displayed strong resistance reaction against all Guatemalan isolates. Even though P. capsici is a cosmopolitan pathogen, this is the first molecular confirmation of this pathogen infecting C. annuum in Guatemala.

Recollections of a Helmstetter Disciple.

Nearly fifty years ago, it became possible to construct E. coli minichromosomes using recombinant DNA technology. These very small replicons, comprising the unique replication origin of the chromosome oriC coupled to a drug resistance marker, provided new opportunities to study the regulation of bacterial chromosome replication, were key to obtaining the nucleotide sequence information encoded into oriC and were essential for the development of a ground-breaking in vitro replication system. However, true authenticity of the minichromosome model system required that they replicate during the cell cycle with chromosome-like timing specificity. I was fortunate enough to have the opportunity to construct E. coli minichromosomes in the laboratory of Charles Helmstetter and, for the first time, measure minichromosome cell cycle regulation. In this review, I discuss the evolution of this project along with some additional studies from that time related to the DNA topology and segregation properties of minichromosomes. Despite the significant passage of time, it is clear that large gaps in our understanding of oriC regulation still remain. I discuss some specific topics that continue to be worthy of further study.

Accelerated Evolution by Diversity-Generating Retroelements.

Diversity-generating retroelements (DGRs) create vast amounts of targeted, functional diversity by facilitating the rapid evolution of ligand-binding protein domains. Thousands of DGRs have been identified in bacteria, archaea, and their respective viruses. They are broadly distributed throughout the microbial world, with enrichment observed in certain taxa and environments. The diversification machinery works through a novel mechanism termed mutagenic retrohoming, whereby nucleotide sequence information is copied from an invariant DNA template repeat (TR) into an RNA intermediate, selectively mutagenized at TR adenines during cDNA synthesis by a DGR-encoded reverse transcriptase, and transferred to a variable repeat (VR) region within a variable-protein gene (54). This unidirectional flow of information leaves TR-DNA sequences unmodified, allowing for repeated rounds of mutagenic retrohoming to optimize variable-protein function. DGR target genes are often modular and can encode one or more of a wide variety of discrete functional domains appended to a diversifiable ligand-binding motif. Bacterial variable proteins often localize to cellsurfaces, although a subset appear to be cytoplasmic, while phage-encoded DGRs commonly diversify tail fiber-associated receptor-binding proteins. Here, we provide a comprehensive review of the mechanism and consequences of accelerated protein evolution by these unique and beneficial genetic elements.

Technological Approaches in the Analysis of Extracellular Vesicle Nucleotide Sequences.

Together with metabolites, proteins, and lipid components, the EV cargo consists of DNA and RNA nucleotide sequence species, which are part of the intracellular communication network regulating specific cellular processes and provoking distinct target cell responses. The extracellular vesicle (EV) nucleotide sequence cargo molecules are often investigated in association with a particular pathology and may provide an insight into the physiological and pathological processes in hard-to-access organs and tissues. The diversity and biological function of EV nucleotide sequences are distinct regarding EV subgroups and differ in tissue- and cell-released EVs. EV DNA is present mainly in apoptotic bodies, while there are different species of EV RNAs in all subgroups of EVs. A limited sample volume of unique human liquid biopsy provides a small amount of EVs with limited isolated DNA and RNA, which can be a challenging factor for EV nucleotide sequence analysis, while the additional difficulty is technical variability of molecular nucleotide detection. Every EV study is challenged with its first step of the EV isolation procedure, which determines the EV’s purity, yield, and diameter range and has an impact on the EV’s downstream analysis with a significant impact on the final result. The gold standard EV isolation procedure with ultracentrifugation provides a low output and not highly pure isolated EVs, while modern techniques increase EV’s yield and purity. Different EV DNA and RNA detection techniques include the PCR procedure for nucleotide sequence replication of the molecules of interest, which can undergo a small-input EV DNA or RNA material. The nucleotide sequence detection approaches with their advantages and disadvantages should be considered to appropriately address the study problem and to extract specific EV nucleotide sequence information with the detection using qPCR or next-generation sequencing. Advanced next-generation sequencing techniques allow the detection of total EV genomic or transcriptomic data even at the single-molecule resolution and thus, offering a sensitive and accurate EV DNA or RNA biomarker detection. Additionally, with the processes where the EV genomic or transcriptomic data profiles are compared to identify characteristic EV differences in specific conditions, novel biomarkers could be discovered. Therefore, a suitable differential expression analysis is crucial to define the EV DNA or RNA differences between conditions under investigation. Further bioinformatics analysis can predict molecular cell targets and identify targeted and affected cellular pathways. The prediction target tools with functional studies are essential to help specify the role of the investigated EV-targeted nucleotide sequences in health and disease and support further development of EV-related therapeutics. This review will discuss the biological diversity of human liquid biopsy–obtained EV nucleotide sequences DNA and RNA species reported as potential biomarkers in health and disease and methodological principles of their detection, from human liquid biopsy EV isolation, EV nucleotide sequence extraction, techniques for their detection, and their cell target prediction.

Molecular identification of lactic acid bacteria an approach to sustainable food security.

BackgroundDadiah is a traditional dish from West Sumatra made from buffalo milk, which is fermented in bamboo tubes and left at room temperature for ±2 days. Dadiah is included in the staple food category because it contains Lactic Acid Bacteria (LAB), which has the potential to be a probiotic. This study aims to determine the identification and characterization of LAB from Dadiah from Halaban, Kab. Fifty Cities, West Sumatra.Design and MethodsA survey method was used in this research with a descriptive analysis, Antimicrobial activity testing was done with bacteria Escherichia coli O157, Staphylococcus aureus, Listeria monocytogenes, and Listeria innocua. Molecular identification was done using the 16S rRNA gene.ResultsProbiotic candidate test with the best results in testing for resistance to stomach acid at pH3 with the viability of 65.98%, bile salt resistance 0.3%, viability of 54.90% from 2DA isolates. Antimicrobial activity with the best clear zone area results was obtained in 2DA isolates with Escherichia coli O157 test bacteria of 21.16 mm, Staphylococcus aureus with a clear zone area of 23.17 mm, Listeria innocua of 19.24 mm and Listeria monocytogenes with a clear zone area 18.23 mm in 4DA isolate, LAB identification using 16S sRNA gene, results of running PCR base length 1419bp.ConclusionsPhylogenetic analysis shows that Dadiah of Limapuluh Kota Regency is a kin to Lactobacillus plantarum. The superiority of identification technology by using 16S rRNA gene only can be conducted if the nucleotide sequence information of the targeted bacteria is known beforehand.

Expression of soluble recombinant human matrix metalloproteinase 9 and generation of its monoclonal antibody

We have successfully produced a recombinant human matrix metalloproteinase 9 (hMMP9) antigen with high yield and purity and used it to generate a hybridoma cell-culture-based monoclonal anti-hMMP9 antibody. We selected the most effective antibody for binding antigens and successfully identified its nucleotide sequence. The entire antigen and antibody developmental procedures described herein can be a practical approach for producing large amounts of monoclonal antibodies against hMMP9 and other antigens of interest. Additionally, the nucleotide sequence information of the anti-hMMP9 monoclonal antibody revealed herein will be useful for the generation of recombinant antibodies or antibody fragments against hMMP9.

Morphology, phylogenetics and pathology of "red sore disease" (coinfection by Epistylis cf. wuhanensis and Aeromonas hydrophila) on sportfishes from reservoirs in the South-Eastern United States.

The aetiological agents of red sore disease (RSD) reportedly comprise a taxonomically ambiguous stalked ciliate (a species of Epistylis) and Aeromonas hydrophila. The taxonomic identity of each pathogen remains provisional: using supra-specific morphological features for the ciliate and culture-based methods that cannot delineate bacterial strain. On 7 and 9 November 2017 and 28 May 2020, biologists and anglers reported a local epizootic (Hiwassee and Chattahoochee river basins; Georgia) wherein some moribund fish presented RSD-like lesions. The ciliates were assigned to Epistylis by morphology. The ciliate is regarded as Epistylis cf wuhanensis, as nucleotide sequences from its small subunit ribosomal DNA were identical to those of Epistylis wuhanensis. The bacterium was identified as Aeromonas hydrophila by phenotypic markers and nucleotide sequences from the DNA gyrase subunit B; our sequences comprised 3 strains and phylogenetically were recovered sister to strains of Eurasian origin. Histological sections of lesions revealed effacement or partial deterioration of the epithelium covering scales, scale loss, haemorrhaging, necrosis, oedema, and extensive inflammatory infiltrate in the dermis. This is the first nucleotide sequence information for the symbionts implicated in RSD.

Intra-strain biological and epidemiological characterization of plum pox virus.

Plum pox virus (PPV) is one of the most important plant viruses causing serious economic losses. Thus far, strain typing based on the definition of 10 monophyletic strains with partially differentiable biological properties has been the sole approach used for epidemiological characterization of PPV. However, elucidating the genetic determinants underlying intra‐strain biological variation among populations or isolates remains a relevant but unexamined aspect of the epidemiology of the virus. In this study, based on complete nucleotide sequence information of 210 Japanese and 47 non‐Japanese isolates of the PPV‐Dideron (D) strain, we identified five positively selected sites in the PPV‐D genome. Among them, molecular studies showed that amino acid substitutions at position 2,635 in viral replicase correlate with viral titre and competitiveness at the systemic level, suggesting that amino acid position 2,635 is involved in aphid transmission efficiency and symptom severity. Estimation of ancestral genome sequences indicated that substitutions at amino acid position 2,635 were reversible and peculiar to one of two genetically distinct PPV‐D populations in Japan. The reversible amino acid evolution probably contributes to the dissemination of the virus population. This study provides the first genomic insight into the evolutionary epidemiology of PPV based on intra‐strain biological variation ascribed to positive selection.

NmSEER V2.0: a prediction tool for 2\u2032-O-methylation sites based on random forest and multi-encoding combination

Background2′-O-methylation (2′-O-me or Nm) is a post-transcriptional RNA methylation modified at 2′-hydroxy, which is common in mRNAs and various non-coding RNAs. Previous studies revealed the significance of Nm in multiple biological processes. With Nm getting more and more attention, a revolutionary technique termed Nm-seq, was developed to profile Nm sites mainly in mRNA with single nucleotide resolution and high sensitivity. In a recent work, supported by the Nm-seq data, we have reported a method in silico for predicting Nm sites, which relies on nucleotide sequence information, and established an online server named NmSEER. More recently, a more confident dataset produced by refined Nm-seq was available. Therefore, in this work, we redesigned the prediction model to achieve a more robust performance on the new data.ResultsWe redesigned the prediction model from two perspectives, including machine learning algorithm and multi-encoding scheme combination. With optimization by 5-fold cross-validation tests and evaluation by independent test respectively, random forest was selected as the most robust algorithm. Meanwhile, one-hot encoding, together with position-specific dinucleotide sequence profile and K-nucleotide frequency encoding were collectively applied to build the final predictor.ConclusionsThe predictor of updated version, named NmSEER V2.0, achieves an accurate prediction performance (AUROC = 0.862) and has been settled into a brand-new server, which is available at http://www.rnanut.net/nmseer-v2/ for free.

Illumina sequencing of clinical samples for virus detection in a public health laboratory

High-throughput sequencing (HTS) provides the opportunity, once a diagnostic result is obtained, to extract additional information from a virus-containing sample. Hence, it offers advantages over established quantitative amplification technology, such as quantitative PCR, particularly in a public health environment. At this early stage of its clinical application, there have been limited studies comparing HTS performance to that of the more established quantitative PCR technology for direct detection of viruses. In this pilot-scale study, we tested HTS with a range of viruses and sample types routinely encountered in a public health virology laboratory. In comparison with quantitative PCR, our HTS method was able to sensitively (92%) detect all viruses in any sample type with the exception of certain tissues. Moreover, sufficient nucleotide sequence information was obtained to enable genotyping of strains detected, thus providing additional useful epidemiological information. While HTS sensitivity may not yet match that of PCR, the added value through enhanced epidemiological data has considerable potential to enable real-time surveillance of circulating strains so as to facilitate rapid and appropriate response to outbreaks and virus zoonotic spillover events.

GBDTCDA: Predicting circRNA-disease Associations Based on Gradient Boosting Decision Tree with Multiple Biological Data Fusion

Circular RNA (circRNA) is a closed-loop structural non-coding RNA molecule which plays a significant role during the gene regulation processes. There are many previous studies shown that circRNAs can be regarded as the sponges of miRNAs. Thus, circRNA is also a key point for disease diagnosing, treating and inferring. However, traditional experimental approaches to verify the associations between the circRNA and disease are time-consuming and money-consuming. There are few computational models to predict potential circRNA-disease associations, which become our motivation to propose a new computational model. In this study, we propose a machine learning based computational model named Gradient Boosting Decision Tree with multiple biological data to predict circRNA-disease associations (GBDTCDA). The known circRNA-disease associations' data are downloaded from cricR2Disease database (http://bioinfo.snnu.edu.cn/CircR2Disease/). The feature vector of each circRNA-disease association pair is composed of four parts, which are the statistics information of different biological networks, the graph theory information of different biological networks, circRNA-disease associations' network information and circRNA nucleotide sequence information, respectively. Therefore, we use those feature vectors to train the gradient boosting decision tree regression model. Then, the leave one out cross validation (LOOCV) is adopted to evaluate the performance of our computational model. As for predicting some common diseases related circRNAs, our method GBDTCDA also obtains the better results. The Area under the ROC Curve (AUC) values of Basal cell carcinoma, Non-small cell lung cancer and cervical cancer are 95.8%, 88.3% and 93.5%, respectively. For further illustrating the performance of GBDTCDA, a case study of breast cancer is also supplemented in this study. Thus, our proposed method GBDTCDA is a powerful tool to predict potential circRNA-disease associations based on experimental results and analyses.

The Challenges of Analysing Highly Diverse Picobirnavirus Sequence Data.

The reliable identification and classification of infectious diseases is critical for understanding their biology and controlling their impact. Recent advances in sequencing technology have allowed insight into the remarkable diversity of the virosphere, of which a large component remains undiscovered. For these emerging or undescribed viruses, the process of classifying unknown sequences is heavily reliant on existing nucleotide sequence information in public databases. However, due to the enormous diversity of viruses, and past focus on the most prevalent and impactful virus types, databases are often incomplete. Picobirnaviridae is a dsRNA virus family with broad host and geographic range, but with relatively little sequence information in public databases. The family contains one genus, Picobirnavirus, which may be associated with gastric illness in humans and animals. Little further information is available due in part to difficulties in identification. Here, we investigate diversity both within the genus Picobirnavirus and among other dsRNA virus types using a combined phylogenetic and functional (protein structure homology-modelling) approach. Our results show that diversity within picobirnavirus exceeds that seen between many other dsRNA genera. Furthermore, we find that commonly used practices employed to classify picobirnavirus, such as analysis of short fragments and trimming of sequences, can influence phylogenetic conclusions. The degree of phylogenetic and functional divergence among picobirnavirus sequences in our study suggests an enormous undiscovered diversity, which contributes to the undescribed “viral dark matter” component of metagenomic studies.

Modelling strategies for assessing and increasing the effectiveness of new phenotyping techniques in plant breeding

New types of phenotyping tools generate large amounts of data on many aspects of plant physiology and morphology with high spatial and temporal resolution. These new phenotyping data are potentially useful to improve understanding and prediction of complex traits, like yield, that are characterized by strong environmental context dependencies, i.e., genotype by environment interactions. For an evaluation of the utility of new phenotyping information, we will look at how this information can be incorporated in different classes of genotype-to-phenotype (G2P) models. G2P models predict phenotypic traits as functions of genotypic and environmental inputs. In the last decade, access to high-density single nucleotide polymorphism markers (SNPs) and sequence information has boosted the development of a class of G2P models called genomic prediction models that predict phenotypes from genome wide marker profiles. The challenge now is to build G2P models that incorporate simultaneously extensive genomic information alongside with new phenotypic information. Beyond the modification of existing G2P models, new G2P paradigms are required. We present candidate G2P models for the integration of genomic and new phenotyping information and illustrate their use in examples. Special attention will be given to the modelling of genotype by environment interactions. The G2P models provide a framework for model based phenotyping and the evaluation of the utility of phenotyping information in the context of breeding programs.

Phylogenetic analysis of the lumpy skin disease viruses in northwest of Iran.

Lumpy skin disease (LSD) is a devastating viral disease of cattle which has recently spread from Africa into the countries of the Middle East. The aim of the present study was to investigate the relationships among lumpy skin disease viruses (LSDV) isolated from different regions of Iran and the origin and spread of these viruses. In this study, a total of 234 blood samples from clinically affected animals from four provinces in the northwest of Iran were screened for LSDV using polymerase chain reaction (PCR). From 80 positive samples for LSDV detected by PCR, the partial P32 gene (759bp) of 12 isolates were sequenced and phylogenetically analyzed. LSD viruses were grouped in three subclusters with an overall 97.1-100% nucleotide identity. LSDVs isolated from Gilan showed lowest nucleotide identity with the other LSDVs. Four isolates of LSDV including KO-1, EA-1, EA-3, and WA-3 showed 100% similarity with each other and also with the Neethling strain. Phylogenetic analysis indicated that the identified LSDVs were closely related to each other and had high-sequence homology with other LSDV isolates from Africa. It was concluded that LSD outbreak probably occurred in the northwest of Iran by LSDVs entering the country from Iraq and P32 nucleotide sequence information obtained in the present study is a valuable resource in understanding the genetic nature and molecular epidemiology of local LSDV isolates which can be used for future vaccine development based on the circulating strains in the region.

The complete mitochondrial genome of a stream loach (Schistura reticulofasciata) and its phylogeny

The stream loach, Schistura reticulofasciata has high demand in ornamental fish trade due to its mesmerizing colorful banding pattern and size. In recent times, its abundance has declined severely in its habitat, and presently it has been assessed as vulnerable fish. Despite its threatened status, no nucleotide sequence information from this fish is publicly available in GenBank and the BOLD System. Here we describe the first complete mitogenome of S. reticulofasciata. The resulting circular mitogenome of S. reticulofasciata is 16,581 in length, comprising 13 protein-coding genes (PCG), 22 transfer RNA (tRNA) genes, 2 ribosomal RNA genes (rRNA), one replication origin (OL) and one control region (D-loop). The overall genome size, gene arrangement, A + T content, AT-skew, GC-skew and partiality towards A and T in mitogenome were similar to those of other Schistura species. Phylogenetic analysis confirmed the high sequence similarity and conserved synteny among the Schistura species. The primary genetic data developed in this study could be successively applied for species identification, genetic diversity studies and the formulation of effective conservation strategies for steam loaches.

A new approach of digital PCR system for non-invasive prenatal screening of trisomy 21

BackgroundNon-invasive prenatal screening (NIPS) of trisomy 21 (T21) using digital PCR (dPCR) with several advantages will be very effective. Here, we developed a dPCR system for T21 screening which allows high sensitivity and real-time diagnosis and thus overcome sequence based analysis. MethodsCut-off value was established using DNA extracted from all 157 T21 negative samples including 47 pregnant woman samples and 3 T21 positive pregnant woman samples extracted from 4 different sample types. To increase the portion of the cell-free fetal DNA (cffDNA) in maternal cell-free DNA (cfDNA), a size selection method was devised. We evaluated the clinical reliability of NIPS using dPCR through analysis of 877 pregnant woman samples. ResultsWe could demonstrate the possibility of NIPS using dPCR performed by applying cut-off value and size selection method. The overall accuracy was derived at 99.66% using 877 pregnant woman plasma samples. ConclusionOur results showed that dPCR can meet the requirements for NIPS for T21. It is relatively inexpensive, easy to use in a screening method and compatible with ethical concerns regarding access to nucleotide sequence information. This study may be a basic data for the development of population-wide screening for T21 in pregnant women.

Don't just dump your data and run: Authors should submit as much experimental information as possible when uploading sequence data.

If you have, in any way, been involved with genetic research over the past 10 years, then you have likely heard of the Sequence Read Archive (SRA), which is jointly housed at the National Center for Biotechnology (NCBI), the DNA Data Bank of Japan (DDBJ), and the European Bioinformatics Institute (EBI). And if you regularly work with genome or transcriptome sequence information, then you have probably extracted data from and/or deposited data into the SRA. For those who are unfamiliar with it, the SRA is an international public online archive for next‐generation sequencing (NGS) data, which was established about a decade ago under the guidance of the International Nucleotide Sequence Database Collaboration (INSDC) [1], [2]. Despite nearly being shut down in 2011 [3], it has grown at a staggering rate over the past 10 years. As of September 1, 2017, it housed over five quadrillion (1015) open‐access bases of NGS data, coming from thousands of different species and spanning the entire gamut of cellular and viral life. It contains DNA‐ and RNA‐sequencing (DNA‐seq and RNA‐seq) reads of every kind, from bisulfite‐seq to strand‐specific RNA‐seq to single‐cell DNA‐seq, and it accepts reads from every type of NGS platform, be it Illumina, Ion Torrent, or PacBio sequencing. In other words, the SRA is a crucial and central resource in the fast‐paced and increasingly important domain of contemporary genetic research. > … the SRA is a crucial and central resource in the fast‐paced and increasingly important domain of contemporary genetic research. ### The Sequence Read Archive The SRA can be easily accessed and searched via the NCBI (https://www.ncbi.nlm.nih.gov/sra), DDBJ (http://trace.ddbj.nig.ac.jp/dra/index_e.html), and EBI (http://www.ebi.ac.uk/ena/submit/read-submission) websites. Once there, you will find yourself at a sequencing‐read superstore. With a decent Wi‐Fi connection, a couple of keyword searches, and a few clicks of the trackpad, you can quickly …

Globalizing Genomics: The Origins of the International Nucleotide Sequence Database Collaboration

Genomics is increasingly considered a global enterprise – the fact that biological information can flow rapidly around the planet is taken to be important to what genomics is and what it can achieve. However, the large-scale international circulation of nucleotide sequence information did not begin with the Human Genome Project. Efforts to formalize and institutionalize the circulation of sequence information emerged concurrently with the development of centralized facilities for collecting that information. That is, the very first databases build for collecting and sharing DNA sequence information were, from their outset, international collaborative enterprises. This paper describes the origins of the International Nucleotide Sequence Database Collaboration between GenBank in the United States, the European Molecular Biology Laboratory Databank, and the DNA Database of Japan. The technical and social groundwork for the international exchange of nucleotide sequences created the conditions of possibility for imagining nucleotide sequences (and subsequently genomes) as a “global” objects. The “transnationalism” of nucleotide sequence was critical to their ontology – what DNA sequences came to be during the Human Genome Project was deeply influenced by international exchange.

De novo assembly and analysis of the Heortia vitessoides transcriptome via high-throughput Illumina sequencing

Heortia vitessoides Moore, a defoliating pest in Aquilaria sinensis (Loureiro) Sprenger forests, is responsible for significant damage to the trees. There is a lack of nucleotide and protein sequence information for H. vitessoides. Hence, in this study, the transcriptome of H. vitessoides was sequenced using the Illumina Hiseq 2000 platform to evaluate the expression profiles and predict the functional genes. A total of 60,587,900 clean reads were produced and de novo assembled into 66,673 unigenes of average length 957bp; 28,202 and 27,339 identified unigenes were annotated using the National Center for Biotechnology Information (NCBI) non-redundant (NR) database and Swiss-Prot database, respectively. All assembled unigenes were categorized into 64 biological processes, cellular components, or molecular functions using Gene Ontology (GO). In total, 24,278 unigenes were annotated using a database of eukaryotic orthologous groups (KOG) and assigned to 25 functional categories. Of these, 9172 unigenes had matches in the Kyoto Encyclopedia of Genes and Genomes (KEGG) and were assigned to 344 KEGG pathways. In addition, there were 11,670 simple sequence repeats (SSRs) in 66,673 unigenes and 64 repeated motif types, of which A/T was the most frequent. We also found that 1000 unigenes were related to insecticide resistance. To the best of our knowledge, this is the first comprehensive transcriptome analysis for H. vitessoides, providing valuable genome data sources for the study of the molecular biology of this pest.