PrimerSNP: a web tool for whole-genome selection of allele-specific and common primers of phylogenetically-related bacterial genomic sequences

Background: The increasing number of genomic sequences of bacteria makes it possible to select unique SNPs of a particular strain/species at the whole genome level and thus design specific primers based on the SNPs. The high similarity of genomic sequences among phylogenetically-related bacteria requires the identification of the few loci in the genome that can serve as unique markers for strain differentiation. PrimerSNP attempts to identify reliable strain-specific markers, on which specific primers are designed for pathogen detection purpose.Results: PrimerSNP is an online tool to design primers based on strain specific SNPs for multiple strains/species of microorganisms at the whole genome level. The allele-specific primers could distinguish query sequences of one strain from other homologous sequences by standard PCR reaction. Additionally, PrimerSNP provides a feature for designing common primers that can amplify all the homologous sequences of multiple strains/species of microorganisms. PrimerSNP is freely available at http://cropdisease.ars.usda.gov/similar to primer.Conclusion: PrimerSNP is a high-throughput specific primer generation tool for the differentiation of phylogenetically-related strains/species. Experimental validation showed that this software had a successful prediction rate of 80.4 - 100% for strain specific primer design.

Conserved sequences in the β subunit of archaeal and eukaryal translation initiation factor 2 (elF2), absent from elF5, mediate interaction with elF2γ

The eukaryotic translation initiation factor 2 (eIF2) binds the methionyl-initiator tRNA in a GTP-dependent mode. This complex associates with the 40 S ribosomal particle, which then, with the aid of other factors, binds to the 5' end of the mRNA and migrates to the first AUG codon, where eIF5 promotes GTP hydrolysis, followed by the formation of the 80 S ribosome. Here we provide a comparative sequence analysis of the β subunit of eIF2 and its archaeal counterpart (aIF2β). aIF2β differs from eIF2β in not possessing an N-terminal extension implicated in binding RNA, eIF5 and eIF2B. The remaining sequences are highly conserved, and are shared with eIF5. Previously isolated mutations in the yeast eIF2β, which allow initiation of translation at UUG codons due to the uncovering of an intrinsic GTPase activity in eIF2, involve residues that are conserved in aIF2β, but not in eIF5. We show that the sequence of eIF2B homologous to aIF2β is sufficient for binding eIF2γ, the only subunit with which it interacts, and comprises, at the most, 78 residues, eIF5 does not interact with eIF2γ, despite its similarity with eIF2β, probably because of a gap in homology in this region. These observations have implications for the evolution of the mechanism of translation initiation.

Molecular and cytogenetic analysis of the telomeric (TTAGGG)n repetitive sequences in the Nile tilapia, Oreochromis niloticus (Teleostei: Cichlidae)

The majority of chromosomes in Oreochromis niloticus, as with most fish karyotyped to date, cannot be individually identified owing to their small size. As a first step in establishing a physical map for this important aquaculture species of tilapia we have analyzed the location of the vertebrate telomeric repeat sequence, (TTAGGG)n, in O. niloticus. Southern blot hybridization analysis and a Bal31 sensitivity assay confirm that the vertebrate telomeric repeat is indeed present at O. niloticus chromosomal ends with repeat tracts extending for 4-10 kb on chromosomal ends in erythrocytes. Fluorescent in situ hybridization revealed that (TTAGGG)n is found not only at telomeres, but also at two interstitial loci on chromosome 1. These data support the hypothesis that chromosome 1, which is significantly larger than all the other chromosomes in the karyotype, was produced by the fusion of three chromosomes and explain the overall reduction of chromosomal number from the ancestral teleost karyotype of 2n=48 to 2n=44 observed in tilapia.

Solutions for a class of integro-differential equations with time periodic coefficients

In this work, a series solution is found for the integro-differential equation y″ (t) = -(ω2 c + ω2 f sin2 ωpt)y(t) + ωf (sin ωpt) z′ (0) + ω2 fωp sin ωpt ∫t 0 (cos ωps) y(s)ds, which describes the charged particle motion for certain configurations of oscillating magnetic fields. As an interesting feature, the terms of the solution are related to distinct sequences of prime numbers.

Phylogenetic relationships among genera Eucalyptus and Corymbia species based on rnDNA internal transcribed spacers sequences

Phylogenetio relationships between Eucalyptus species, subgenus Symphyomyrtus (sections Adnataria, Exsertaria, Maldenaria, and Transversaria), and Corymbia species (sections Politaria and Ocharia) were established based on the sequence of Internal transcribed rDNA spacers (ITS1 and ITS2). The species analyzed were obtained from a collection kept in Brazil. Fragments obtained using primers ITS1 and ITS2 were sequenced and part of the sequence of ITS1 and ITS2 and the complete sequence of 5.8S rDNA were used in the analysis. ITSs and 5.8S rDNA sequences from E. globulus ssp. globulus and A. bakeri (Genus Angophora) were downloaded from the Genbank database and included in the analysis. Psidlum guajava was the selected outgroup used. The sequence alignment and a Neighbor-joining tree were obtained using Clustal X. Few variations were detected in the 5.8S rDNA sequences obtained, occurring mainly between Eucalyptus and Corymbia, thus defining these genera. Variations in ITS sequences occurred in all investigated species. Phylogenetic analysis showed a clear separation between the genera Corymbia and Eucalyptus. A bakeri was more closely related to species belonging to genus Corymbia. Regarding the subgenus Symphyomyrtus (Genus Eucalyptus), only species from section Maidenaria grouped together according to their common section. This could have been caused by the removal of natural reproductive barriers when these species were introduced In Brazil, with a consequent Increase in the rate of interspecific crossings and Introgression events.

Comparative analysis of noncoding sequences of orthologous bovine and human gene pairs

Genomic sequence comparison across species has enabled the elucidation of important coding and regulatory sequences encoded within DNA. Of particular interest are the noncoding regulatory sequences, which influence gene transcriptional and posttranscriptional processes. A phylogenetic footprinting strategy was employed to identify noncoding conservation patterns of 39 human and bovine orthologous genes. Seventy-three conserved noncoding sequences were identified that shared greater than 70% identity over at least 100 bp. Thirteen of these conserved sequences were also identified in the mouse genome. Evolutionary conservation of noncoding sequences across diverse species may have functional significance, and these conserved sequences may be good candidates for regulatory elements.

Identification and complete sequencing of novel human transcripts through the use of mouse orthologs and testis cDNA sequences

The correct identification of all human genes, and their derived transcripts, has not yet been achieved, and it remains one of the major aims of the worldwide genomics community. Computational programs suggest the existence of 30,000 to 40,000 human genes. However, definitive gene identification can only be achieved by experimental approaches. We used two distinct methodologies, one based on the alignment of mouse orthologous sequences to the human genome, and another based on the construction of a high-quality human testis cDNA library, in an attempt to identify new human transcripts within the human genome sequence. We generated 47 complete human transcript sequences, comprising 27 unannotated and 20 annotated sequences. Eight of these transcripts are variants of previously known genes. These transcripts were characterized according to size, number of exons, and chromosomal localization, and a search for protein domains was undertaken based on their putative open reading frames. In silico expression analysis suggests that some of these transcripts are expressed at low levels and in a restricted set of tissues.

Phylogenetic analysis of flatfish (order pleuronectiformes) based on mitochondrial 16s rDNA sequences

The phylogenetic relationships of the order Pleuronectiformes are controversial and at some crucial points remain unresolved. To date most phylogenetic studies on this order have been based on morpho-anatomical criteria, whereas only a few sequence comparisons based studies have been reported. In the present study, the phylogenetic relationships of 30 flatfish species pertaining to seven different families were examined by sequence analysis of the first half of the 16S mitochondrial DNA gene. The results obtained did not support percoids as the sister group of pleuronectiforms. The monophyletic origin of most families analyzed, Soleidae, Scophthalmidae, Achiridae, Pleuronectidae and Bothidae, was strongly supported, except for Paralichthyidae which was clearly subdivided into two groups, one of them associated with high confidence to Pleuronectidae. The analysis of the 16S rRNA gene also suggested the monophyly of Pleuronectiforms as the most probable hypothesis and consistently supported some major interfamily groupings.

Taxonomic and phylogenetic relationships between neotropical species of ticks from genus Amblyomma (Acari: Ixodidae) inferred from second internal transcribed spacer sequences of rDNA

The accurate specific identification of ticks is essential for the study, control and prevention of tick-borne diseases. Herein, we determined ribosomal nucleotide sequences of the second internal transcribed spacer (ITS2) of 15 Neotropical hard tick species of the genus Amblyomma Koch found in Brazil. Most of the studied ticks accidentally parasite humans and potentially act as vectors of zoonoses. Lengths of the ITS2 sequences ranged from 956 to 1,207 bp, whereas GC content varied from 62.4 to 66.9%. A matrix of ITS2 divergence was calculated with the ITS2 sequence data obtained showing divergence levels varying from 1.5 to 28.8%. The analysis indicated that this molecular marker can be useful for Amblyomma-specific identification. Phylogenetic inferences based on the ITS2 sequences were used to assess some issues in subgenus taxonomy. © 2007 Entomological Society of America.

Nuclear mitochondrial-like sequences in ants: Evidence from Atta cephalotes (Formicidae: Attini)

Nuclear mitochondrial-like sequences (numts) are copies of mitochondrial DNA that have migrated to the genomic DNA. We present the first characterization of numts in ants, these numts being homologues to a mitochondrial DNA fragment containing loci the 3′ portion of the cytochrome oxidase I gene, an intergenic spacer, the tRNA leucine gene and the 5′ portion of the cytochrome oxidase II gene. All 67 specimens of Atta cephalotes (Hymenoptera: Formicidae: Attini) investigated had these homologues, which are within two monophyletic groups that we called numt1 and numt2. Numt1 and numt2 sequences are less variable than mitochondrial sequences and released from the severe purifying selection constraining the evolution of mitochondrial genes. Their formation probably involved bottlenecks related to two distinct transfer events of ancient and fast evolving mitochondrial DNA fragments to comparative slowly evolving nuclear DNA regions. © 2007 The Authors.

Weak mixing and eigenvalues for arnoux-rauzy sequences

We define by simple conditions two wide subclasses of the socalled Arnoux-Rauzy systems; the elements of the first one share the property of (measure-theoretic) weak mixing, thus we generalize and improve a counterexample to the conjecture that these systems are codings of rotations; those of the second one have eigenvalues, which was known hitherto only for a very small set of examples.

Potential scenarios for Internet use in the mathematics classroom

Research on the influence of multiple representations in mathematics education gained new momentum when personal computers and software started to become available in the mid-1980s. It became much easier for students who were not fond of algebraic representations to work with concepts such as function using graphs or tables. Research on how students use such software showed that they shaped the tools to their own needs, resulting in an intershaping relationship in which tools shape the way students know at the same time the students shape the tools and influence the design of the next generation of tools. This kind of research led to the theoretical perspective presented in this paper: knowledge is constructed by collectives of humans-with-media. In this paper, I will discuss how media have shaped the notions of problem and knowledge, and a parallel will be developed between the way that software has brought new possibilities to mathematics education and the changes that the Internet may bring to mathematics education. This paper is, therefore, a discussion about the future of mathematics education. Potential scenarios for the future of mathematics education, if the Internet becomes accepted in the classroom, will be discussed. © FIZ Karlsruhe 2009.

Collectives of humans-with-media in mathematics education: Notebooks, blackboards, calculators, computers and ... notebooks throughout 100 years of ICMI

The main aim of this study was to present evidence of the ways in which different media have conditioned and dramatically reorganized education, in general, and mathematics education, in particular. After an introduction of the theme, we discuss the epistemological perspective that provides the foundation for our analysis: the notion of humans-with-media. Then, we briefly illustrate how the medium is related to the scientific production of mathematical knowledge. We take a detour into the world of art to examine how devices and instruments have historically been associated with the production of mathematical knowledge. Then, we review studies on the history of education to show how traditional media were introduced into schools and have influenced education. In particular, we examine how devices such as blackboards and notebooks, which were novelties a 100 years ago, came to be accepted in schools and the mathematical activities that were promoted with their use. Finally, we discuss how information technology has changed education and how the Internet may have an impact on mathematics education comparable to that of the notebook over a century ago. © FIZ Karlsruhe 2009.