RESUMO
Plasmids are extrachromosomal DNA elements of microorganisms encoding beneficial genetic information. They were thought to be equally distributed to daughter cells during cell division. Here we use mathematical modeling to investigate the evolutionary stability of plasmid segregation for high-copy plasmids-plasmids that are present in up to several hundred copies per cell-carrying antibiotic resistance genes. Evolutionary stable strategies (ESS) are determined by numerical analysis of a plasmid-load structured population model. The theory predicts that the evolutionary stable segregation strategy of a cell depends on the plasmid copy number: For low and medium plasmid load, both daughters receive in average an equal share of plasmids, while in case of high plasmid load, one daughter obtains distinctively and systematically more plasmids. These findings are in good agreement with recent experimental results. We discuss the interpretation and practical consequences.
Assuntos
Evolução Biológica , Modelos Biológicos , Plasmídeos , Resistência Microbiana a Medicamentos/genéticaRESUMO
Different biomolecules have been identified in bacterial pathogens that sense changes in temperature and trigger expression of virulence programs upon host entry. However, the dynamics and quantitative outcome of this response in individual cells of a population, and how this influences pathogenicity are unknown. Here, we address these questions using a thermosensing virulence regulator of an intestinal pathogen (RovA of Yersinia pseudotuberculosis) as a model. We reveal that this regulator is part of a novel thermoresponsive bistable switch, which leads to high- and low-invasive subpopulations within a narrow temperature range. The temperature range in which bistability is observed is defined by the degradation and synthesis rate of the regulator, and is further adjustable via a nutrient-responsive regulator. The thermoresponsive switch is also characterized by a hysteretic behavior in which activation and deactivation occurred on vastly different time scales. Mathematical modeling accurately mirrored the experimental behavior and predicted that the thermoresponsiveness of this sophisticated bistable switch is mainly determined by the thermo-triggered increase of RovA proteolysis. We further observed RovA ON and OFF subpopulations of Y. pseudotuberculosis in the Peyer's patches and caecum of infected mice, and that changes in the RovA ON/OFF cell ratio reduce tissue colonization and overall virulence. This points to a bet-hedging strategy in which the thermoresponsive bistable switch plays a key role in adapting the bacteria to the fluctuating conditions encountered as they pass through the host's intestinal epithelium and suggests novel strategies for the development of antimicrobial therapies.
Assuntos
Proteínas de Bactérias/metabolismo , Fatores de Transcrição/metabolismo , Fatores de Virulência/metabolismo , Infecções por Yersinia pseudotuberculosis/parasitologia , Yersinia pseudotuberculosis/patogenicidade , Animais , Western Blotting , Modelos Animais de Doenças , Ensaio de Desvio de Mobilidade Eletroforética , Feminino , Citometria de Fluxo , Camundongos , Camundongos Endogâmicos BALB C , Temperatura , Imagem com Lapso de Tempo , VirulênciaRESUMO
During the past 2 decades, Bacillus megaterium has been systematically developed for the gram-per-liter scale production of recombinant proteins. The plasmid-based expression systems employed use a xylose-controlled promoter. Protein production analyses at the single-cell level using green fluorescent protein as a model product revealed cell culture heterogeneity characterized by a significant proportion of less productive bacteria. Due to the enormous size of B. megaterium, such bistable behavior seen in subpopulations was readily analyzed by time lapse microscopy and flow cytometry. Cell culture heterogeneity was not caused simply by plasmid loss: instead, an asymmetric distribution of plasmids during cell division was detected during the exponential-growth phase. Multicopy plasmids are generally randomly distributed between daughter cells. However, in vivo and in vitro experiments demonstrated that under conditions of strong protein production, plasmids are retained at one of the cell poles. Furthermore, it was found that cells with accumulated plasmids and high protein production ceased cell division. As a consequence, the overall protein production of the culture was achieved mainly by the subpopulation with a sufficient plasmid copy number. Based on our experimental data, we propose a model whereby the distribution of multicopy plasmids is controlled by polar fixation under protein production conditions. Thereby, cell lines with fluctuating plasmid abundance arise, which results in population heterogeneity. Our results provide initial insights into the mechanism of cellular heterogeneity during plasmid-based recombinant protein production in a Bacillus species.
Assuntos
Bacillus megaterium/citologia , Bacillus megaterium/metabolismo , Polaridade Celular , Proteínas de Fluorescência Verde/metabolismo , Plasmídeos/genética , Proteínas Recombinantes/metabolismo , Bacillus megaterium/genética , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Proteínas de Fluorescência Verde/genética , Plasmídeos/metabolismo , Proteínas Recombinantes/genéticaRESUMO
Efficient adaptation strategies to changing environmental conditions are essential for bacteria to survive and grow. Fundamental restructuring of their metabolism is usually mediated by corresponding gene regulation. Here, often several different environmental stimuli have to be integrated into a reasonable, energy-efficient response. Fast fluctuations and overshooting have to be filtered out. The gene regulatory network for the anaerobic adaptation of the pathogenic bacterium Pseudomonas aeruginosa is organized as a feed-forward loop (FFL), which is a three-gene network motif composed of two transcription factors (Anr for oxygen, NarxL for nitrate) and one target (Nar for nitrate reductase). The upstream transcription factor (Anr) induces the downstream transcription factor (NarXL). Both regulators act together positively by inducing the target (Nar) via a direct and indirect regulation path (coherent type-1 FFL). Since full promoter activity is only achieved when both transcription factors are present the target operon is expressed with a delay. Thus, in response to environmental stimuli (oxygen, nitrate), signals are mediated and processed in a way that short pulses are filtered out. In this study we analyze a special kind of FFL called FFLk by means of a family of ordinary differential equation models. The secondary FFL regulator (NarXL) is expressed constitutively but further induced in the presence of the upstream stimuli. This FFL modification has substantial influence on the response time and cost-benefit ratio mediated by environmental fluctuations. In order to find conditions where this regulatory network motif might be beneficial, we analyzed various models and environments. We describe the observed evolutional advantage of FFLk and its role in environmental adaptation and pathogenicity.
Assuntos
Adaptação Biológica/fisiologia , Meio Ambiente , Redes Reguladoras de Genes/fisiologia , Modelos Biológicos , Pseudomonas aeruginosa/fisiologia , Adaptação Biológica/genética , Anaerobiose , Análise Custo-Benefício , Pseudomonas aeruginosa/genéticaRESUMO
BACKGROUND: The knowledge of metabolic pathways and fluxes is important to understand the adaptation of organisms to their biotic and abiotic environment. The specific distribution of stable isotope labelled precursors into metabolic products can be taken as fingerprints of the metabolic events and dynamics through the metabolic networks. An open-source software is required that easily and rapidly calculates from mass spectra of labelled metabolites, derivatives and their fragments global isotope excess and isotopomer distribution. RESULTS: The open-source software "Least Square Mass Isotopomer Analyzer" (LS-MIDA) is presented that processes experimental mass spectrometry (MS) data on the basis of metabolite information such as the number of atoms in the compound, mass to charge ratio (m/e or m/z) values of the compounds and fragments under study, and the experimental relative MS intensities reflecting the enrichments of isotopomers in 13C- or 15 N-labelled compounds, in comparison to the natural abundances in the unlabelled molecules. The software uses Brauman's least square method of linear regression. As a result, global isotope enrichments of the metabolite or fragment under study and the molar abundances of each isotopomer are obtained and displayed. CONCLUSIONS: The new software provides an open-source platform that easily and rapidly converts experimental MS patterns of labelled metabolites into isotopomer enrichments that are the basis for subsequent observation-driven analysis of pathways and fluxes, as well as for model-driven metabolic flux calculations.
Assuntos
Espectrometria de Massas/métodos , Redes e Vias Metabólicas , Software , Algoritmos , Isótopos , Análise dos Mínimos Quadrados , Modelos BiológicosRESUMO
MOTIVATION: Time-lapse imaging in combination with fluorescence microscopy techniques enable the investigation of gene regulatory circuits and uncovered phenomena like culture heterogeneity. In this context, computational image processing for the analysis of single cell behaviour plays an increasing role in systems biology and mathematical modelling approaches. Consequently, we developed a software package with graphical user interface for the analysis of single bacterial cell behaviour. RESULTS: A new software called TLM-Tracker allows for the flexible and user-friendly interpretation for the segmentation, tracking and lineage analysis of microbial cells in time-lapse movies. AVAILABILITY: The software package, including manual, tutorial video and examples, is available as Matlab code or executable binaries at http://www.tlmtracker.tu-bs.de.
Assuntos
Processamento de Imagem Assistida por Computador/métodos , Microscopia de Fluorescência/métodos , Microscopia de Vídeo/métodos , Análise de Célula Única/métodos , Software , Imagem com Lapso de Tempo/métodos , Bacillus megaterium/citologia , Bacillus megaterium/metabolismo , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Fluorescência Verde/química , Modelos Biológicos , Modelos TeóricosRESUMO
MOTIVATION: InFiRe, Insertion Finder via Restriction digest, is a novel software tool that allows for the computational identification of transposon insertion sites in known bacterial genome sequences after transposon mutagenesis experiments. The approach is based on the fact that restriction endonuclease digestions of bacterial DNA yield a unique pattern of DNA fragments with defined sizes. Transposon insertion changes the size of the hosting DNA fragment by a known number of base pairs. The exact size of this fragment can be determined by Southern blot hybridization. Subsequently, the position of insertion can be identified with computational analysis. The outlined method provides a solid basis for the establishment of a new high-throughput technology. AVAILABILITY AND IMPLEMENTATION: The software is freely available on our web server at www.infire.tu-bs.de. The algorithm was implemented in the statistical programming language R. For the most flexible use, InFiRe is provided in two different versions. A web interface offers the convenient use in a web browser. In addition, the software and source code is freely available for download as R-packages on our website. CONTACT: m.steinert@tu-bs.de SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.
Assuntos
Algoritmos , Mutagênese Insercional , Elementos de DNA Transponíveis , DNA Bacteriano/genética , Genoma Bacteriano , Internet , Legionella pneumophila/genética , SoftwareRESUMO
Streptococcus pyogenes (group A streptococcus [GAS]) is a highly virulent Gram-positive bacterium. For successful infection, GAS expresses many virulence factors, which are clustered together with transcriptional regulators in distinct genomic regions. Ralp3 is a central regulator of the ERES region. In this study, we investigated the role of Ralp3 in GAS M49 pathogenesis. The inactivation of Ralp3 resulted in reduced attachment to and internalization into human keratinocytes. The Δralp3 mutant failed to survive in human blood and serum, and the hyaluronic acid capsule was slightly decreased. In addition, the mutant showed a lower binding capacity to human plasminogen, and the SpeB activity was significantly decreased. Complementation of the Δralp3 mutant restored the wild-type phenotype. The transcriptome and quantitative reverse transcription-PCR analysis of the serotype M49 GAS strain and its isogenic Δralp3 mutant identified 16 genes as upregulated, and 43 genes were found to be downregulated. Among the downregulated genes, there were open reading frames encoding proteins involved in metabolism (e.g., both lac operons and the fru operon), genes encoding lantibiotics (e.g., the putative salivaricin operon), and ORFs encoding virulence factors (such as the whole Mga core regulon and further genes under Mga control). In summary, the ERES region regulator Ralp3 is an important serotype-specific transcriptional regulator for virulence and metabolic control.
Assuntos
Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica/fisiologia , Streptococcus pyogenes/metabolismo , Streptococcus pyogenes/patogenicidade , Fatores de Virulência/metabolismo , Aderência Bacteriana , Proteínas de Bactérias/genética , Linhagem Celular Tumoral , Perfilação da Expressão Gênica , Humanos , Ácido Hialurônico , Mutação , Fenótipo , RNA Bacteriano , Transcriptoma , Virulência , Fatores de Virulência/genéticaRESUMO
UNLABELLED: GeneReporter is a web tool that reports functional information and relevant literature on a protein-coding sequence of interest. Its purpose is to support both manual genome annotation and document retrieval. PubMed references corresponding to a sequence are detected by the extraction of query words from UniProt entries of homologous sequences. Data on protein families, domains, potential cofactors, structure, function, cellular localization, metabolic contribution and corresponding DNA binding sites complement the information on a given gene product of interest. AVAILABILITY AND IMPLEMENTATION: GeneReporter is available at http://www.genereporter.tu-bs.de. The web site integrates databases and analysis tools as SOAP-based web services from the EBI (European Bioinformatics Institute) and NCBI (National Center for Biotechnology Information).
Assuntos
Anotação de Sequência Molecular , Análise de Sequência de Proteína , Software , Biologia Computacional , Mineração de Dados , Bases de Dados Genéticas , Internet , Proteínas/química , Proteínas/genética , PubMed , Homologia de Sequência de AminoácidosRESUMO
Analysis of the genome sequence of the small hyperthermophilic archaeal parasite Nanoarchaeum equitans has not revealed genes encoding the glutamate, histidine, tryptophan and initiator methionine transfer RNA species. Here we develop a computational approach to genome analysis that searches for widely separated genes encoding tRNA halves that, on the basis of structural prediction, could form intact tRNA molecules. A search of the N. equitans genome reveals nine genes that encode tRNA halves; together they account for the missing tRNA genes. The tRNA sequences are split after the anticodon-adjacent position 37, the normal location of tRNA introns. The terminal sequences can be accommodated in an intervening sequence that includes a 12-14-nucleotide GC-rich RNA duplex between the end of the 5' tRNA half and the beginning of the 3' tRNA half. Reverse transcriptase polymerase chain reaction and aminoacylation experiments of N. equitans tRNA demonstrated maturation to full-size tRNA and acceptor activity of the tRNA(His) and tRNA(Glu) species predicted in silico. As the joining mechanism possibly involves tRNA trans-splicing, the presence of an intron might have been required for early tRNA synthesis.
Assuntos
Genes Arqueais/genética , Nanoarchaeota/genética , RNA Arqueal/genética , RNA de Transferência/genética , Trans-Splicing/genética , Aminoacilação , Sequência de Bases , Biologia Computacional , Genoma Arqueal , Genômica , Glutamato-tRNA Ligase/metabolismo , Histidina-tRNA Ligase/metabolismo , Íntrons/genética , Dados de Sequência Molecular , Nanoarchaeota/enzimologia , Conformação de Ácido Nucleico , RNA Arqueal/química , RNA Arqueal/isolamento & purificação , RNA Arqueal/metabolismo , RNA de Transferência/química , RNA de Transferência/isolamento & purificação , RNA de Transferência/metabolismoRESUMO
Streptococci are the causative agent of many human infectious diseases including bacterial pneumonia and meningitis. Here, we present Strepto-DB, a database for the comparative genome analysis of group A (GAS) and group B (GBS) streptococci. The known genomes of various GAS and GBS contain a large fraction of distributed genes that were found absent in other strains or serotypes of the same species. Strepto-DB identifies the homologous proteins deduced from the genomes of interest. It allows for the elucidation of the GAS and GBS core- and pan-genomes via genome-wide comparisons. Moreover, an intergenic region analysis tool provides alignments and predictions for transcription factor binding sites in the non-coding sequences. An interactive genome browser visualizes functional annotations. Strepto-DB (http://oger.tu-bs.de/strepto_db) was created by the use of OGeR, the Open Genome Resource for comparative analysis of prokaryotic genomes. OGeR is a newly developed open source database and tool platform for the web-based storage, distribution, visualization and comparison of prokaryotic genome data. The system automatically creates the dedicated relational database and web interface and imports an arbitrary number of genomes derived from standardized genome files. OGeR can be downloaded at http://oger.tu-bs.de.
Assuntos
Bases de Dados Genéticas , Genoma Bacteriano , Streptococcus/genética , Proteínas de Bactérias/genética , DNA Intergênico/química , Genômica , Streptococcus/classificaçãoRESUMO
PRODORIC is a database that provides annotated information on the regulation of gene expression in prokaryotes. It integrates a large compilation of gene regulatory data including transcription factor binding sites, promoter structures and gene expression patterns. The whole dataset is manually curated and relies on published results extracted from the scientific literature. The current extended version of PRODORIC contains gene regulatory data for several new microorganisms. Major improvements were realized in the design of the web interface and the accessibility of the stored information. The database was further improved by the implementation of various new tools for the elucidation of gene regulatory interactions. Thus, the PRODORIC platform represents a framework for the interactive exploration, prediction and evaluation of gene regulatory networks in prokaryotes. PRODORIC is accessible at http://www.prodoric.de.
Assuntos
Bases de Dados Genéticas , Regulação Bacteriana da Expressão Gênica , Software , Sítios de Ligação , Redes Reguladoras de Genes , Regiões Promotoras Genéticas , Fatores de Transcrição/metabolismoRESUMO
The anaerobic metabolism of the opportunistic pathogen Pseudomonas aeruginosa is important for growth and biofilm formation during persistent infections. The two Fnr-type transcription factors Anr and Dnr regulate different parts of the underlying network in response to oxygen tension and NO. Little is known about all members of the Anr and Dnr regulons and the mediated immediate response to oxygen depletion. Comprehensive transcriptome and bioinformatics analyses in combination with a limited proteome analyses were used for the investigation of the P. aeruginosa response to an immediate oxygen depletion and for definition of the corresponding Anr and Dnr regulons. We observed at first the activation of fermentative pathways for immediate energy generation followed by induction of alternative respiratory chains. A solid position weight matrix model was deduced from the experimentally identified Anr boxes and used for identification of 170 putative Anr boxes in potential P. aeruginosa promoter regions. The combination with the experimental data unambiguously identified 130 new members for the Anr and Dnr regulons. The basis for the understanding of two regulons of P. aeruginosa central to biofilm formation and infection is now defined.
Assuntos
Adaptação Fisiológica , Proteínas de Bactérias/metabolismo , Pseudomonas aeruginosa/fisiologia , Regulon , Transativadores/metabolismo , Fatores de Transcrição/metabolismo , Anaerobiose , Proteínas de Bactérias/genética , Fermentação , Análise em Microsséries , Dados de Sequência Molecular , Nitrato Redutase/metabolismo , Óxido Nítrico/metabolismo , Oxigênio/metabolismo , Transativadores/genética , Fatores de Transcrição/genéticaRESUMO
ProdoNet is a web-based application for the mapping of prokaryotic genes and the corresponding proteins to common gene regulatory and metabolic networks. For a given list of genes, the system detects shared operons, identifies co-expressed genes and deduces joint regulators. In addition, the contribution to shared metabolic pathways becomes visible on KEGG maps. Furthermore, the co-occurrence of genes of interest in gene expression profiles can be added to the visualization of the global network. In this way, ProdoNet provides the basis for functional genomics approaches and for the interpretation of transcriptomics and proteomics data. As an example, we present an investigation of an experimental membrane subproteome analysis of Pseudomonas aeruginosa with ProdoNet. The ProdoNet dataset on transcriptional regulation is based on the PRODORIC Prokaryotic Database of Gene Regulation and the Virtual Footprint tool. ProdoNet is accessible at http://www.prodonet.tu-bs.de.
Assuntos
Bactérias/metabolismo , Redes Reguladoras de Genes , Genes Bacterianos , Software , Proteínas de Bactérias/genética , Gráficos por Computador , Perfilação da Expressão Gênica , Internet , Proteínas de Membrana/genética , Redes e Vias Metabólicas , Óperon , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/metabolismo , Regulon , Interface Usuário-ComputadorRESUMO
To provide an integrated bioinformatics platform for a systems biology approach to the biology of pseudomonads in infection and biotechnology the database SYSTOMONAS (SYSTems biology of pseudOMONAS) was established. Besides our own experimental metabolome, proteome and transcriptome data, various additional predictions of cellular processes, such as gene-regulatory networks were stored. Reconstruction of metabolic networks in SYSTOMONAS was achieved via comparative genomics. Broad data integration is realized using SOAP interfaces for the well established databases BRENDA, KEGG and PRODORIC. Several tools for the analysis of stored data and for the visualization of the corresponding results are provided, enabling a quick understanding of metabolic pathways, genomic arrangements or promoter structures of interest. The focus of SYSTOMONAS is on pseudomonads and in particular Pseudomonas aeruginosa, an opportunistic human pathogen. With this database we would like to encourage the Pseudomonas community to elucidate cellular processes of interest using an integrated systems biology strategy. The database is accessible at http://www.systomonas.de.
Assuntos
Bases de Dados Genéticas , Pseudomonas/genética , Biologia de Sistemas , Proteínas de Bactérias/classificação , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Sistemas de Gerenciamento de Base de Dados , Redes Reguladoras de Genes , Genoma Bacteriano , Genômica , Internet , Redes e Vias Metabólicas , Pseudomonas/metabolismo , Infecções por Pseudomonas/microbiologia , Integração de Sistemas , Interface Usuário-ComputadorRESUMO
A novel program suite was implemented for the functional interpretation of high-throughput gene expression data based on the identification of Gene Ontology (GO) nodes. The focus of the analysis lies on the interpretation of microarray data from prokaryotes. The three well established statistical methods of the threshold value-based Fisher's exact test, as well as the threshold value-independent Kolmogorov-Smirnov and Student's t-test were employed in order to identify the groups of genes with a significantly altered expression profile. Furthermore, we provide the application of the rank-based unpaired Wilcoxon's test for a GO-based microarray data interpretation. Further features of the program include recognition of the alternative gene names and the correction for multiple testing. Obtained results are visualized interactively both as a table and as a GO subgraph including all significant nodes. Currently, JProGO enables the analysis of microarray data from more than 20 different prokaryotic species, including all important model organisms, and thus constitutes a useful web service for the microbial research community. JProGO is freely accessible via the web at the following address: http://www.jprogo.de.
Assuntos
Perfilação da Expressão Gênica/métodos , Genes Arqueais , Genes Bacterianos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Software , Archaea/genética , Bactérias/genética , Gráficos por Computador , Interpretação Estatística de Dados , Internet , Interface Usuário-Computador , Vocabulário ControladoRESUMO
MOTIVATION: After the publication of JVirGel 1.0 in 2003 we got many requests and suggestions from the proteomics community to further improve the performance of the software and to add additional useful new features. RESULTS: The integration of the PrediSi algorithm for the prediction of signal peptides for the Sec-dependent protein export into JVirGel 2.0 allows the exclusion of most exported preproteins from calculated proteomic maps and provides the basis for the calculation of Sec-based secretomes. A tool for the identification of transmembrane helices carrying proteins (JCaMelix) and the prediction of the corresponding membrane proteome was added. Finally, in order to directly compare experimental and calculated proteome data, a function to overlay and evaluate predicted and experimental two-dimensional gels was included. AVAILABILITY: JVirGel 2.0 is freely available as precompiled package for the installation on Windows or Linux operating systems. Furthermore, there is a completely platform-independent Java version available for download. Additionally, we provide a Java Server Pages based version of JVirGel 2.0 which can be operated in nearly all web browsers. All versions are accessible at http://www.jvirgel.de
Assuntos
Eletroforese em Gel Bidimensional/métodos , Proteínas de Membrana/química , Mapeamento de Peptídeos/métodos , Proteoma/química , Proteoma/metabolismo , Análise de Sequência de Proteína/métodos , Software , Algoritmos , Sequência de Aminoácidos , Proteínas de Membrana/análise , Dados de Sequência Molecular , Sinais Direcionadores de Proteínas , Proteoma/análise , Alinhamento de Sequência/métodosRESUMO
UNLABELLED: MetaQuant is a Java-based program for the automatic and accurate quantification of GC/MS-based metabolome data. In contrast to other programs MetaQuant is able to quantify hundreds of substances simultaneously with minimal manual intervention. The integration of a self-acting calibration function allows the parallel and fast calibration for several metabolites simultaneously. Finally, MetaQuant is able to import GC/MS data in the common NetCDF format and to export the results of the quantification into Systems Biology Markup Language (SBML), Comma Separated Values (CSV) or Microsoft Excel (XLS) format. AVAILABILITY: MetaQuant is written in Java and is available under an open source license. Precompiled packages for the installation on Windows or Linux operating systems are freely available for download. The source code as well as the installation packages are available at http://bioinformatics.org/metaquant
Assuntos
Inteligência Artificial , Cromatografia Gasosa-Espectrometria de Massas/métodos , Perfilação da Expressão Gênica/métodos , Mapeamento de Peptídeos/métodos , Proteoma/química , Proteoma/metabolismo , Software , Algoritmos , Interface Usuário-ComputadorRESUMO
The database VBASE2 provides germ-line sequences of human and mouse immunoglobulin variable (V) genes. It acts as an interconnecting platform between several existing self-contained data systems: VBASE2 integrates genome sequence data and links to the V genes in the Ensembl Genome Browser. For a single V gene sequence, all references to the EMBL nucleotide sequence database are provided, including references for V(D)J rearrangements. Furthermore, cross-references to the VBASE database, the IMGT database and the Kabat database are available. A DAS server allows the display of VBASE2 V genes within the Ensembl Genome Browser. VBASE2 can be accessed either by a web-based text query or by a sequence similarity search with the DNAPLOT software. VBASE2 is available at http://www.vbase2.org, and the DAS server is located at http://www.dnaplot.com/das.
Assuntos
Bases de Dados de Ácidos Nucleicos , Genes de Imunoglobulinas , Região Variável de Imunoglobulina/genética , Animais , Humanos , Camundongos , Integração de Sistemas , Interface Usuário-ComputadorRESUMO
A novel method for the adaptation of target gene codon usage to most sequenced prokaryotes and selected eukaryotic gene expression hosts was developed to improve heterologous protein production. In contrast to existing tools, JCat (Java Codon Adaptation Tool) does not require the manual definition of highly expressed genes and is, therefore, a very rapid and easy method. Further options of JCat for codon adaptation include the avoidance of unwanted cleavage sites for restriction enzymes and Rho-independent transcription terminators. The output of JCat is both graphically and as Codon Adaptation Index (CAI) values given for the pasted sequence and the newly adapted sequence. Additionally, a list of genes in FASTA-format can be uploaded to calculate CAI values. In one example, all genes of the genome of Caenorhabditis elegans were adapted to Escherichia coli codon usage and further optimized to avoid commonly used restriction sites. In a second example, the Pseudomonas aeruginosa exbD gene codon usage was adapted to E.coli codon usage with parallel avoidance of the same restriction sites. For both, the degree of introduced changes was documented and evaluated. JCat is integrated into the PRODORIC database that hosts all required information on the various organisms to fulfill the requested calculations. JCat is freely accessible at http://www.prodoric.de/JCat.