ZFIN, the Zebrafish Model Organism Database, http://zfin.org, serves as the central repository and web-based resource for zebrafish genetic, genomic, phenotypic and developmental data. ZFIN manually curates comprehensive data for zebrafish genes, phenotypes, genotypes, gene expression, antibodies, anatomical structures and publications. A wide-ranging collection of web-based search forms and tools facilitates access to integrated views of these data promoting analysis and scientific discovery. Data represented in ZFIN are derived from three primary sources: curation of zebrafish publications, individual research laboratories and collaborations with bioinformatics organizations. Data formats include text, images and graphical representations. ZFIN is a dynamic resource with data added daily as part of our ongoing curation process. Software updates are frequent. Here, we describe recent additions to ZFIN including (i) enhanced access to images, (ii) genomic features, (iii) genome browser, (iv) transcripts, (v) antibodies and (vi) a community wiki for protocols and antibodies.

ABSTRACT10 The Zebrafish Information Network (ZFIN; http://zfin. org) is a web based community resource that implements the curation of zebrafish genetic, genomic and developmental data. ZFIN provides an integrated representation of mutants, genes, genetic markers, 15 mapping panels, publications and community resources such as meeting announcements and contact information. Recent enhancements to ZFIN include (i) comprehensive curation of gene expression data from the literature and from directly sub-20 mitted data, (ii) increased support and annotation of the genome sequence, (iii) expanded use of ontologies to support curation and query forms, (iv) curation of morpholino data from the literature, and (v) increased versatility of gene pages, with new data 25 types, links and analysis tools.

The Zebrafish Information Network (ZFIN, http://zfin.org), the model organism database for zebrafish, provides the central location for curated zebrafish genetic, genomic and developmental data. Extensive data integration of mutant phenotypes, genes, expression patterns, sequences, genetic markers, morpholinos, map positions, publications and community resources facilitates the use of the zebrafish as a model for studying gene function, development, behavior and disease. Access to ZFIN data is provided via web-based query forms and through bulk data files. ZFIN is the definitive source for zebrafish gene and allele nomenclature, the zebrafish anatomical ontology (AO) and for zebrafish gene ontology (GO) annotations. ZFIN plays an active role in the development of cross-species ontologies such as the phenotypic quality ontology (PATO) and the gene ontology (GO). Recent enhancements to ZFIN include (i) a new home page and navigation bar, (ii) expanded support for genotypes and phenotypes, (iii) comprehensive phenotype annotations based on anatomical, phenotypic quality and gene ontologies, (iv) a BLAST server tightly integrated with the ZFIN database via ZFIN-specific datasets, (v) a global site search and (vi) help with hands-on resources.

The Zebrafish Information Network (ZFIN) is a web based community resource that serves as a centralized location for the curation and integration of zebrafish genetic, genomic and developmental data. ZFIN is publicly accessible at http://zfin.org. ZFIN provides an integrated representation of mutants, genes, genetic markers, mapping panels, publications and community contact data. Recent enhancements to ZFIN include: (i) an anatomical dictionary that provides a controlled vocabulary of anatomical terms, grouped by developmental stages, that may be used to annotate and query gene expression data; (ii) gene expression data; (iii) expanded support for genome sequence; (iv) gene annotation using the standardized vocabulary of Gene Ontology (GO) terms that can be used to elucidate relationships between gene products in zebrafish and other organisms; and (v) collaborations with other databases (NCBI, Sanger Institute and SWISS-PROT) to provide standardization and interconnections based on shared curation.

The Zebrafish Information Network, ZFIN, is a WWW community resource of zebrafish genetic, genomic and developmental research information (http://zfin.org). ZFIN provides an anatomical atlas and dictionary, developmental staging criteria, research methods, pathology information and a link to the ZFIN relational database (http://zfin. org/ZFIN/). The database, built on a relational, object-oriented model, provides integrated information about mutants, genes, genetic markers, mapping panels, publications and contact information for the zebrafish research community. The database is populated with curated published data, user submitted data and large dataset uploads. A broad range of data types including text, images, graphical representations and genetic maps supports the data. ZFIN incorporates links to other genomic resources that provide sequence and ortholog data. Zebrafish nomenclature guidelines and an automated registration mechanism for new names are provided. Extensive usability testing has resulted in an easy to learn and use forms interface with complex searching capabilities.

Using a family of synthetic tetradecamer oligonucleotides as a primer for cDNA synthesis and a second family of tetradecamers as a hybridization probe, we have prepared and isolated a cDNA clone of mouse myelin basic protein (MBP). The clone, pNZ111, corresponds to the region of the mRNA that codes for an amino acid sequence present in all four major forms of MBP (2)(3)(4)(5). Large amounts of the membrane are produced relative to the size of the myelin-forming cell. The process involves the synthesis of myelin-specific proteins followed by their integration into the growing membrane. During early myelinogenesis, membranes are formed that are loosely whorled around the axon. As myelin maturation proceeds, compaction of these membranes occurs to form the tight multilamellar structure characteristic of myelin.The myelin basic protein (MBP) constitutes about 33% of the protein of the myelin sheath in the central nervous system and 1-10% in the peripheral (6, 7). In most species, the predominant form of the MBP has a molecular mass of 18.5 kilodaltons (kDa), and the primary sequence of this protein is well conserved in animals as distantly related as the chicken, human, and rat (8-10). Two major forms of MBP predominate in rats and mice: one of these corresponds to the single major protein found in other species (18.5 kDa) and the other has a molecular mass of 14 kDa. In rats, the 14-kDa MBP is identical in sequence to the 18.5-kDa MBP, except for an internal deletion of residues 118-158 [numbered according to Martenson (11)] near the carboxyl terminus of the protein (10, 12). Two quantitatively minor forms of MBP have been found in rats and mice-a 21.5-kDa protein that is presumably identical to the 18.5-kDa MBP, except for the inclusion of a 25-30 amino acid sequence in the NH2-terminal half of the molecule and a 17-kDa MBP that bears the same relationship to the 14-kDa MBP (13).Myelination occurs postnatally in mouse brain beginning 8-10 days postpartum and continuing actively for 7-10 wk, with the maximal rate of myelin deposition occurring at about 18 days (4). Maximal synthesis of the 18.5-kDa and 14-kDa MBPs occurs at 18 days in vivo and coincides closely with the peak of myelin synthesis (14). During myelin maturation in the mouse, the proportions of the four MBPs in the membrane change (15-17) and this may be due to alterations in the relative rates of synthesis of the four proteins with age. With maturation, the proportion of the two minor MBPs (i.e., the 21.5 kDa and 17 kDa in myelin falls relative to the 18.5-kDa proteins and the 14-kDa/18.5-kDa MBP ratio increases dramatically. This latter change has been correlated with the relative rate of synthesis of these two proteins in vivo (14). Recently, Carson et al. (18) have identified trace amounts of larger forms of MBP that appear during the early stages of myelination. Identification of these larger forms has depended on immunoreagents and the relationships of the amino acid sequences to the major MBPs have, as yet, not been determined.We have r...

A cDNA clone containing the entire vesicular stomatitis virus nucleocapsid gene was assembled by fusing portions of two partial clones. When the cDNA clone was inserted into a new general-purpose eucaryotic expression vector and introduced into appropriate host cells, abundant N-protein synthesis ensued. The expressed protein was indistinguishable from authentic N protein produced during vesicular stomatitis virus infections. The recombinant N protein was recognized by a polyclonal antibody and two different monoclonal antibodies and could not be resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from authentic N. Our results suggest that the recombinant N protein produced in transfected cells rapidly aggregates into high-molecular-weight complexes in the absence of vesicular stomatitis virus genomic RNA.

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