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RIKEN mouse genome encyclopedia.

We have been working to establish the comprehensive mouse full-length cDNA collection and sequence database to cover as many genes as we can, named Riken mouse genome encyclopedia. Recently we are constructing higher-level annotation (Functional ANnoTation Of Mouse cDNA; FANTOM) not only with homology search based annotation but also with expression data profile, mapping information and protein-protein database. More than 1,000,000 clones prepared from 163 tissues were end-sequenced to classify into 159,789 clusters and 60,770 representative clones were fully sequenced. As a conclusion, the 60,770 sequences contained 33,409 unique. The next generation of life science is clearly based on all of the genome information and resources. Based on our cDNA clones we developed the additional system to explore gene function. We developed cDNA microarray system to print all of these cDNA clones, protein-protein interaction screening system, protein-DNA interaction screening system and so on. The integrated database of all the information is very useful not only for analysis of gene transcriptional network and for the connection of gene to phenotype to facilitate positional candidate approach. In this talk, the prospect of the application of these genome resourced should be discussed. More information is available at the web page: http://genome.gsc.riken.go.jp/.

Animals↗

SpliceInfo: an information repository for mRNA alternative splicing in human genome.

We have developed an information repository named SpliceInfo to collect the occurrences of the four major alternative-splicing (AS) modes in human genome; these include exon skipping, 5'-alternative splicing, 3'-alternative splicing and intron retention. The dataset is derived by comparing the nucleotide and protein sequences available for a given gene for evidence of AS. Additional features such as the tissue specificity of the mRNA, the protein domain contained by exons, the GC-ratio of exons, the repeats contained within the exons, and the Gene Ontology are annotated computationally for each exonic region that is alternatively spliced. Motivated by a previous investigation of AS-related motifs such as exonic splicing enhancer and exonic splicing silencer, this resource also provides a means of identifying motifs candidates and this should help to identify potential regulatory mechanisms within a particular exonic sequence set and its two flanking intronic sequence sets. This is carried out using motif discovery tools to identify motif candidates related to alternative splicing regulation and together with a secondary structure prediction tool, will help in the identification of the structural properties of such regulatory motifs. The integrated resource is now available on http://SpliceInfo.mbc.NCTU.edu.tw/.

Alternative Splicing↗

The Schmidtea mediterranea database as a molecular resource for studying platyhelminthes, stem cells and regeneration.

Platyhelminthes are excellent models for the study of stem cell biology, regeneration and the regulation of scale and proportion. In addition, parasitic forms infect millions of people worldwide. Therefore, it is puzzling that they remain relatively unexplored at the molecular level. We present the characterization of approximately 3,000 non-redundant cDNAs from a clonal line of the planarian Schmidtea mediterranea. The obtained cDNA sequences, homology comparisons and high-throughput whole-mount in situ hybridization data form part of the S. mediterranea database (SmedDb; http://planaria.neuro.utah.edu). Sixty-nine percent of the cDNAs analyzed share similarities with sequences deposited in GenBank and dbEST. The remaining gene transcripts failed to match sequences in other organisms, even though a large number of these (approximately 80%) contained putative open reading frames. Taken together, the molecular resources presented in this study, along with the ability of abrogating gene expression in planarians using RNA interference technology, pave the way for a systematic study of the remarkable biological properties displayed by Platyhelminthes.

Animals↗

Fusarium genomic resources: tools to limit crop diseases and mycotoxin contamination.

It has been almost 10 years since Joan Bennett suggested that fungal biologists create a "wish list" for fungal genome sequences (Bennett JW. White paper: Genomics for filamentous fungi. Fungal Genet Biol 1997; 21: 3-7). The availability of over 200 review papers concerning fungal genomics is a reflection of significant progress with a diversity of fungal species. Although much progress has been made, the use of genomic data to study mycotoxin synthesis and function, pathogenesis and other aspects of fungal biology is in its infancy. Here, we briefly present the status of publicly available genomic resources for Fusarium, a genus of important plant pathogenic and mycotoxin-producing fungi of worldwide concern. Preliminary examination of microarray data collected from F. verticillioides liquid cultures provides evidence of widespread differential gene expression over time.

Crops, Agricultural↗

Microarray expression profiling resources for plant genomics.

Large volumes of genomic data have been generated for several plant species over the past decade, including structural sequence data and functional annotation at the genome level. Various technologies such as expressed sequence tags (ESTs), massively parallel signature sequencing (MPSS) and microarrays have been used to study gene expression and to provide functional data for many genes simultaneously. This review focuses on recent advances in the application of microarrays in plant genomic research and in gene expression databases available for plants. Large sets of Arabidopsis microarray data are publicly available. Recently developed array platforms are currently being used to generate genome-wide expression profiles for several crop species. Coupled to these platforms are public databases that provide access to these large-scale expression data, which can be used to aid the functional discovery of gene function.

Arabidopsis↗

Advances in spotted microarray resources for expression profiling.

The new millennium has ushered in a new era in human biology. The elucidation of the human genome sequence, together with those of model organisms, provides us with an unprecedented insight into the makeup of our genetic blueprint. The challenge now is to figure out how all the constituent pieces fit together to form the whole picture, and the consequences of what happens when the process goes awry. One experimental tool that has the potential to provide enormous insights into this complex process is expression profiling using microarrays. The past few years have seen a considerable growth in the availability and use of microarrays. Fuelled in part by the many genome projects currently underway, there has been a large increase in the number of organisms for which microarray reagents are available from both commercial and academic sources. In addition to the increasing number of genome-wide probe sets that are available, a significant amount of attention has been focussed on generating more targeted probe sets that focus in on specific pathways or biological processes. Finally, the microarray field is starting to see a shift away from the use of cDNAs or polymerase chain reaction (PCR) products as probes towards the use of 50-70mer oligonucleotide probes with all of the potential advantages that they offer. The aim of this review is to provide an overview of what is currently available in terms of spotted microarray reagents both with respect to pre-made arrays and to probe sets available for arraying.

Computational Biology↗

Development and characterization of genetic mapping resources for the turkey (Meleagris gallopavo).

The development and partial characterization of turkey genomic libraries enriched for TG, GAT, and CCT simple sequence repeats (SSR) are described. The primary library, established using conventional methods, was enriched for each of the three SSR by single-primer polymerase chain reaction (PCR). The three enriched libraries were screened by standard hybridization and washing protocols under moderate to high stringency conditions. The utility of a fraction of the markers was evaluated based on the polymorphism of PCR-amplified products in a backcross reference DNA panel. The panel consisted of genomic DNA samples from three backcrossed families developed from a cross of a wild male turkey to three inbred Orlopp line C females. A total of 181 sequences from positive clones have been characterized and deposited in GenBank. About 60% of the 60 primer pairs designed from SSR-containing sequences detected polymorphism in the reference DNA panel. The turkey genomic DNA reference panel, the enriched libraries, and the markers described here provide an opportunity to begin to characterize the turkey genome and to develop a useful public genetic map for this economically important species.

Animals↗

Gene homology resources on the World Wide Web.

As the amount of information available to biologists increases exponentially, data analysis becomes progressively more challenging. Sequence homology has been a traditional tool in the researchers' armamentarium; it is a very versatile instrument and can be employed to assist in numerous tasks, from establishing the function of a gene to determination of the evolutionary development of an organism. Consequently, numerous specialized tools have been established in the public domain (most commonly, the World Wide Web) to help investigators use sequence homology in their research. These homology databases differ both in techniques they use to compare sequences as well as in the size of the unit of analysis, which can be the whole gene, a domain, or a motif. In this paper, we aim to present a systematic review of the inner details of the most commonly used databases as well as to offer guidelines for their use.

Animals↗

HmtDB, a human mitochondrial genomic resource based on variability studies supporting population genetics and biomedical research.

BACKGROUND: Population genetics studies based on the analysis of mtDNA and mitochondrial disease studies have produced a huge quantity of sequence data and related information. These data are at present worldwide distributed in differently organised databases and web sites not well integrated among them. Moreover it is not generally possible for the user to submit and contemporarily analyse its own data comparing them with the content of a given database, both for population genetics and mitochondrial disease data. RESULTS: HmtDB is a well-integrated web-based human mitochondrial bioinformatic resource aimed at supporting population genetics and mitochondrial disease studies, thanks to a new approach based on site-specific nucleotide and aminoacid variability estimation. HmtDB consists of a database of Human Mitochondrial Genomes, annotated with population data, and a set of bioinformatic tools, able to produce site-specific variability data and to automatically characterize newly sequenced human mitochondrial genomes. A query system for the retrieval of genomes and a web submission tool for the annotation of new genomes have been designed and will soon be implemented. The first release contains 1255 fully annotated human mitochondrial genomes. Nucleotide site-specific variability data and multialigned genomes can be downloaded. Intra-human and inter-species aminoacid variability data estimated on the 13 coding for proteins genes of the 1255 human genomes and 60 mammalian species are also available. HmtDB is freely available, upon registration, at http://www.hmdb.uniba.it. CONCLUSION: The HmtDB project will contribute towards completing and/or refining haplogroup classification and revealing the real pathogenic potential of mitochondrial mutations, on the basis of variability estimation.

Computational Biology↗

PRINTS and PRINTS-S shed light on protein ancestry.

The PRINTS database houses a collection of protein fingerprints. These may be used to make family and tentative functional assignments for uncharacterised sequences. The September 2001 release (version 32.0) includes 1600 fingerprints, encoding approximately 10 000 motifs, covering a range of globular and membrane proteins, modular polypeptides and so on. In addition to its continued steady growth, we report here its use as a source of annotation in the InterPro resource, and the use of its relational cousin, PRINTS-S, to model relationships between families, including those beyond the reach of conventional sequence analysis approaches. The database is accessible for BLAST, fingerprint and text searches at http://www.bioinf.man.ac.uk/dbbrowser/PRINTS/.

Amino Acid Motifs↗

NAST: a multiple sequence alignment server for comparative analysis of 16S rRNA genes.

Microbiologists conducting surveys of bacterial and archaeal diversity often require comparative alignments of thousands of 16S rRNA genes collected from a sample. The computational resources and bioinformatics expertise required to construct such an alignment has inhibited high-throughput analysis. It was hypothesized that an online tool could be developed to efficiently align thousands of 16S rRNA genes via the NAST (Nearest Alignment Space Termination) algorithm for creating multiple sequence alignments (MSA). The tool was implemented with a web-interface at http://greengenes.lbl.gov/NAST. Each user-submitted sequence is compared with Greengenes' 'Core Set', comprising approximately 10,000 aligned non-chimeric sequences representative of the currently recognized diversity among bacteria and archaea. User sequences are oriented and paired with their closest match in the Core Set to serve as a template for inserting gap characters. Non-16S data (sequence from vector or surrounding genomic regions) are conveniently removed in the returned alignment. From the resulting MSA, distance matrices can be calculated for diversity estimates and organisms can be classified by taxonomy. The ability to align and categorize large sequence sets using a simple interface has enabled researchers with various experience levels to obtain bacterial and archaeal community profiles.

Algorithms↗

PhyloNaP: a user-friendly database of phylogeny for natural product-producing enzymes.

SUMMARY: Phylogenetic analysis is widely used to predict enzyme function, yet building annotated and reusable trees is labor-intensive and requires extensive knowledge about the specific enzymes. Existing resources rarely cover biosynthetic enzymes and lack the context needed for meaningful analysis. We present PhyloNaP, the first large-scale resource dedicated to phylogenies of biosynthetic enzymes. PhyloNaP provides ∼51 000 annotated and interactive trees enriched with chemical, functional, and taxonomic information. Users can classify their own sequences via phylogenetic placement, enabling functional inference in an evolutionary context. A contribution portal allows the community to submit curated trees. By combining scale, breadth of annotation, and interactive functionality, PhyloNaP fills a major gap in bioinformatics resources for enzyme discovery and annotation, with immediate applications to secondary metabolism and beyond. AVAILABILITY AND IMPLEMENTATION: Freely available on the web at https://phylonap.cs.uni-tuebingen.de.

Phylogeny↗

The European Bioinformatics Institute's data resources.

As the amount of biological data grows, so does the need for biologists to store and access this information in central repositories in a free and unambiguous manner. The European Bioinformatics Institute (EBI) hosts six core databases, which store information on DNA sequences (EMBL-Bank), protein sequences (SWISS-PROT and TrEMBL), protein structure (MSD), whole genomes (Ensembl) and gene expression (ArrayExpress). But just as a cell would be useless if it couldn't transcribe DNA or translate RNA, our resources would be compromised if each existed in isolation. We have therefore developed a range of tools that not only facilitate the deposition and retrieval of biological information, but also allow users to carry out searches that reflect the interconnectedness of biological information. The EBI's databases and tools are all available on our website at www.ebi.ac.uk.

Animals↗

SIMAP--the similarity matrix of proteins.

MOTIVATION: Sequence similarity searches are of great importance in bioinformatics. Exhaustive searches for homologous proteins in databases are computationally expensive and can be replaced by a database of pre-calculated homologies in many cases. Retrieving similarities from an incrementally updated database instead of repeatedly recalculating them should provide homologs much faster and frees computational resources for other purposes. RESULTS: We have implemented SIMAP-a database containing the similarity space formed by almost all amino acid sequences from public databases and completely sequenced genomes. The database is capable of handling very large datasets and allows incremental updates. We have implemented a powerful backbone for similarity computation, which is based on FASTA heuristics. By providing WWW interfaces as well as web services, we make our data accessible to the worldwide community. We have also adapted procedures to detect putative orthologs as example applications. AVAILABILITY: The SIMAP portal page providing links to SIMAP services is publicly available: http://mips.gsf.de/services/analysis/simap/. The web services can be accessed under http://mips.gsf.de/proj/hobitws/services/RPCSimapService?wsdl and http://mips.gsf.de/proj/hobitws/services/DocSimapService?wsdl

Algorithms↗

Application of fluorescence-based resistance gene analog analysis for genotyping plant genetic resources.

Conventional approaches for detecting disease resistance gene analogs (RGAs) in plants are based on agarose gels or on polyacrylamide gel electrophoresis (PAGE) in combination with silver staining or radioactive labeling. A modified method for RGA analysis has been developed by using fluorescence-labeled primers for PCR amplifications. The amplified fragments are detected by denaturing PAGE using an automated laser fluorescence DNA sequencer and analyzed by fragment analysis software. This technique is not limited to specific plant species and is suitable for high-throughput genotyping plant genetic resources. We demonstrate here the efficiency of this method for comparison of RGA patterns in diverse plant species and for genotyping of natural populations of the wheat progenitor, Triticum dicoccoides.

Electrophoresis, Gel, Two-Dimensional↗

Ovarian gene database.

OBJECTIVE: The entire human genome will be sequenced in September 2000. Facing the exponential increase of data in the GenBank at the National Center for Biotechnology Information, reproductive biologists are being bombarded with massive amounts of information on diverse genes. It is becoming increasingly difficult for individual investigators to sort out the diverse genetic and physiologic information on the localization and function of different genes in the ovary. To alleviate the present situation, we have taken advantage of the accessibility of the Internet and initiated a project that serves the entire ovarian research community. RESULTS AND DISCUSSION: The Ovarian Kaleidoscope database provides information regarding biologic function, expression pattern, and regulation of genes that are expressed in the ovary. In addition, it serves as a gateway to other online information resources relevant to ovarian research by offering results from original papers and data about nucleotide and amino acid sequences, and human and murine mutation phenotypes. All references are linked by hypertext to PubMed and additional links to sequence databases are also included. This information is accessible online and searchable not only by gene name but also by criteria such as the cellular and ovarian function of the gene product, the expression of genes in different ovarian cell types, or their association with specific ovarian phenotypes.

Base Sequence↗

The role of pattern databases in sequence analysis.

In the wake of the numerous now-fruitful genome projects, we are entering an era rich in biological data. The field of bioinformatics is poised to exploit this information in increasingly powerful ways, but the abundance and growing complexity both of the data and of the tools and resources required to analyse them are threatening to overwhelm us. Databases and their search tools are now an essential part of the research environment. However, the rate of sequence generation and the haphazard proliferation of databases have made it difficult to keep pace with developments. In an age of information overload, researchers want rapid, easy-to-use, reliable tools for functional characterisation of newly determined sequences. But what are those tools? How do we access them? Which should we use? This review focuses on a particular type of database that is increasingly used in the task of routine sequence analysis--the so-called pattern database. The paper aims to provide an overview of the current status of pattern databases in common use, outlining the methods behind them and giving pointers on their diagnostic strengths and weaknesses.

Amino Acid Motifs↗