949 research outputs found
Stable nuclear transformation of Eudorina elegans
No full textLerche K, Hallmann A. Stable nuclear transformation of Eudorina elegans. BMC Biotechnology. 2013;13(1): 11.UNLABELLED: ABSTRACT: BACKGROUND: A fundamental step in evolution was the transition from unicellular to differentiated, multicellular organisms. Volvocine algae have been used for several decades as a model lineage to investigate the evolutionary aspects of multicellularity and cellular differentiation. There are two well-studied volvocine species, a unicellular alga (Chlamydomonas reinhardtii) and a multicellular alga with differentiated cell types (Volvox carteri). Species with intermediate characteristics also exist, which blur the boundaries between unicellularity and differentiated multicellularity. These species include the globular alga Eudorina elegans, which is composed of 16-32 cells. However, detailed molecular analyses of E. elegans require genetic manipulation. Unfortunately, genetic engineering has not yet been established for Eudorina, and only limited DNA and/or protein sequence information is available. RESULTS: Here, we describe the stable nuclear transformation of E. elegans by particle bombardment using both a chimeric selectable marker and reporter genes from different heterologous sources. Transgenic algae resistant to paromomycin were achieved using the aminoglycoside 3'-phosphotransferase VIII (aphVIII) gene of Streptomyces rimosus, an actinobacterium, under the control of an artificial promoter consisting of two V. carteri promoters in tandem. Transformants exhibited an increase in resistance to paromomycin by up to 333-fold. Co-transformation with non-selectable plasmids was achieved with a rate of 50 - 100%. The luciferase (gluc) gene from the marine copepod Gaussia princeps, which previously was engineered to match the codon usage of C. reinhardtii, was used as a reporter gene. The expression of gluc was mediated by promoters from C. reinhardtii and V. carteri. Heterologous heat shock promoters induced an increase in luciferase activity (up to 600-fold) at elevated temperatures. Long-term stability and both constitutive and inducible expression of the co-bombarded gluc gene was demonstrated by transcription analysis and bioluminescence assays. CONCLUSIONS: Heterologous flanking sequences, including promoters, work in E. elegans and permit both constitutive and inducible expression of heterologous genes. Stable nuclear transformation of E. elegans is now routine. Thus, we show that genetic engineering of a species is possible even without the resources of endogenous genes and promoters
There is more than one way to turn a spherical cellular monolayer inside out: type B embryo inversion in Volvox globator
No full textHöhn S, Hallmann A. There is more than one way to turn a spherical cellular monolayer inside out: type B embryo inversion in Volvox globator. BMC Biology. 2011;9(1): 89.Background:
Epithelial folding is a common morphogenetic process during the development of multicellular organisms. In metazoans, the biological and biomechanical processes that underlie such three-dimensional (3D) developmental events are usually complex and difficult to investigate. Spheroidal green algae of the genus Volvox are uniquely suited as model systems for studying the basic principles of epithelial folding. Volvox embryos begin life inside out and then must turn their spherical cell monolayer outside in to achieve their adult configuration; this process is called 'inversion.' There are two fundamentally different sequences of inversion processes in Volvocaceae: type A and type B. Type A inversion is well studied, but not much is known about type B inversion. How does the embryo of a typical type B inverter, V. globator, turn itself inside out?
Results:
In this study, we investigated the type B inversion of V. globator embryos and focused on the major movement patterns of the cellular monolayer, cell shape changes and changes in the localization of cytoplasmic bridges (CBs) connecting the cells. Isolated intact, sectioned and fragmented embryos were analyzed throughout the inversion process using light microscopy, confocal laser scanning microscopy, scanning electron microscopy and transmission electron microscopy techniques. We generated 3D models of the identified cell shapes, including the localizations of CBs. We show how concerted cell-shape changes and concerted changes in the position of cells relative to the CB system cause cell layer movements and turn the spherical cell monolayer inside out. The type B inversion of V. globator is compared to the type A inversion in V. carteri.
Conclusions:
Concerted, spatially and temporally coordinated changes in cellular shapes in conjunction with concerted migration of cells relative to the CB system are the causes of type B inversion in V. globator. Despite significant similarities between type A and type B inverters, differences exist in almost all details of the inversion process, suggesting analogous inversion processes that arose through parallel evolution. Based on our results and due to the cellular biomechanical implications of the involved tensile and compressive forces, we developed a global mechanistic scenario that predicts epithelial folding during embryonic inversion in V. globator
Stable nuclear transformation of Gonium pectorale
No full textLerche K, Hallmann A. Stable nuclear transformation of Gonium pectorale. BMC Biotechnology. 2009;9(1): 64.BACKGROUND: Green algae of the family Volvocaceae are a model lineage for studying the molecular evolution of multicellularity and cellular differentiation. The volvocine alga Gonium is intermediate in organizational complexity between its unicellular relative, Chlamydomonas, and its multicellular relatives with differentiated cell types, such as Volvox. Gonium pectorale consists of ~16 biflagellate cells arranged in a flat plate. The detailed molecular analysis of any species necessitates its accessibility to genetic manipulation, but, in volvocine algae, transformation procedures have so far only been established for Chlamydomonas reinhardtii and Volvox carteri. RESULTS: Stable nuclear transformation of G. pectorale was achieved using a heterologous dominant antibiotic resistance gene, the aminoglycoside 3'-phosphotransferase VIII gene (aphVIII) of Streptomyces rimosus, as a selectable marker. Heterologous 3'- and 5'-untranslated flanking sequences, including promoters, were from Chlamydomonas reinhardtii or from Volvox carteri. After particle gun bombardment of wild type Gonium cells with plasmid-coated gold particles, transformants were recovered. The transformants were able to grow in the presence of the antibiotic paromomycin and produced a detectable level of the AphVIII protein. The plasmids integrated into the genome, and stable integration was verified after propagation for over 1400 colony generations. Co-transformants were recovered with a frequency of ~30-50% when cells were co-bombarded with aphVIII-based selectable marker plasmids along with unselectable plasmids containing heterologous genes. The transcription of the co-transformed, unselectable genes was confirmed. After heterologous expression of the luciferase gene from the marine copepod Gaussia princeps, which was previously engineered to match the codon usage in C. reinhardtii, Gonium transformants show luciferase activity through light emission in bioluminescence assays. CONCLUSION: Flanking sequences that include promoters from C. reinhardtii and from V. carteri work in G. pectorale and allow the functional expression of heterologous genes, such as the selectable marker gene aphVIII of S. rimosus or the co-transformed, codon-optimized G. princeps luciferase gene, which turned out to be a suitable reporter gene in Gonium. The availability of a method for transformation of Gonium makes genetic engineering of this species possible and allows for detailed studies in molecular evolution using the unicellular Chlamydomonas, the 16-celled Gonium, and the multicellular Volvox
A new perspective on exploring the Cooper/Eromanga petroleum province - evidence of oil charging from the Warburton Basin
No full textC. O. E. Hallmann, K. R. Arouri, D. M. McKirdy and L. Schwar
Algal and fungal diversity on various dimension stone substrata in the Saale/Unstrut region
No full textConsumer profiles of women’s football spectators
No full textHallmann K, Giel T, Beermann S, Herold E, Breuer C. Consumer profiles of women’s football spectators. Soccer & Society. 2021:1-15
Assessing customer segments and drivers of involvement among ice hockey attendees
No full textHallmann K, Wicker P, Breuer C, Rumpf C, Bode M. Assessing customer segments and drivers of involvement among ice hockey attendees. International Journal of Sport Management. 2013;14:61-80
Quantitative analysis of cell-type specific gene expression in the green alga Volvox carteri
No full textNematollahi G, Kianianmomeni A, Hallmann A. Quantitative analysis of cell-type specific gene expression in the green alga Volvox carteri. BMC Genomics. 2006;7(1): 321.Background: The multicellular alga Volvox carteri possesses only two cell types: mortal, motile somatic cells and potentially immortal, immotile reproductive cells. It is therefore an attractive model system for studying how cell-autonomous cytodifferentiation is programmed within a genome. Moreover, there are ongoing genome projects both in Volvox carteri and in the closely related unicellular alga Chlamydomonas reinhardtii. However, gene sequencing is only the beginning. To identify cell-type specific expression and to determine relative expression rates, we evaluate the potential of real-time RT-PCR for quantifying gene transcript levels. Results: Here we analyze a diversified pool of 39 target genes by real-time RT-PCR for each cell type. This gene pool contains previously known genes with unknown localization of cellular expression, 28 novel genes which are described in this study for the first time, and a few known, cell-type specific genes as a control. The respective gene products are, for instance, part of photosynthesis, cellular regulation, stress response, or transport processes. We provide expression data for all these genes. Conclusion: The results show that quantitative real-time RT-PCR is a favorable approach to analyze cell-type specific gene expression in Volvox, which can be extended to a much larger number of genes or to developmental or metabolic mutants. Our expression data also provide a basis for a detailed analysis of individual, previously unknown, cell-type specifically expressed genes
The influence of microbial mats on travertine precipitation in active hydrothermal systems (Central Italy)
No full textThe study of hydrothermal travertines contributes to the understanding of the interaction between physico-chemical processes and microbial mats in carbonate precipitation. Three active travertine sites were investigated in Central Italy to characterise the types of carbonate precipitates and the associated microbial mats at varying physico-chemical parameters. Carbonate precipitated fabrics at the decimetre to millimetre-scale and microbial mat composition vary with decreasing water temperature: (a) at relatively higher temperature (55–44°C) calcite and aragonite crystals precipitate on microbial mats of Chloroflexi and sulphur-oxidizing microbes forming filamentous streamer fabrics with sparse cyanobacteria, (b) at intermediate temperature (44–40°C), rafts, coated gas bubbles and dendrites are associated with Spirulina cyanobacteria and other filamentous and rod-shaped cyanobacteria, (c) low temperature (34–33°C) laminated crusts and oncoids forming in a terraced slope system are associated with diverse Oscillatoriales and Nostocales filamentous cyanobacteria, Spirulina and diatoms. At the microscale, carbonate precipitates are similar in the three sites consisting of prismatic calcite crystals organised in radial rosettes or fibrous aragonite spherulites (40–300 μm in diameter), overlying or embedded in Extracellular Polymeric Substances. Clotted peloidal micrite dominates at temperatures <40°C, also encrusting filamentous microbes. Carbonates are associated with gypsum crystals; extracellular polymeric substances are enriched in silicon, aluminium, magnesium, calcium, phosphorous and sulphur; authigenic aluminium-silicates form aggregates on Extracellular Polymeric Substances. This study confirms that microbial communities in hydrothermal settings vary as a function of water temperature. Carbonate precipitates at the microscale are similar in the three settings, despite different microbial communities, suggesting that travertine precipitation, driven by carbon dioxide degassing, is influenced by biofilm extracellular polymeric substances acting as a substrate for crystal nucleation (Extracellular Polymeric Substances-mediated mineralization) and affecting the resultant fabric types, independently from specific microbial community composition and metabolism
Financing of sport facilities in Germany
No full textBreuer C, Hallmann K, Wicker P, Feiler S. Financing of sport facilities in Germany. In: Chaix P, ed. Les grands stades. Au cœur des enjeux économiques et sociaux entre collectivités publiques et clubs professionnels. La librairie des humanités . Paris: L'Harmattan; 2011: 135-152
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