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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    World journal of microbiology and biotechnology 15 (1999), S. 411-415 
    ISSN: 1573-0972
    Keywords: Electroporation ; Micrococcus species ; steroid biotransformation ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A steroid-biotransforming strain RJ6 was identified as Micrococcus roseus. This bacterium has a 10 kb plasmid pMQV10. Curing mediated through cultivation of the culture with a low concentration (200 ng/ml) of mitomycin C is described. Loss of cholesterol degradation (chol+) and streptomycin resistance (Smr) phenotypes as a consequence of the loss of plasmid indicate the extrachromosomal location of these two genes in this strain. An electroporation procedure was developed for transformation of cured strain of Micrococcus (RJC6) by plasmids. Frequency of greater than 105 transformants/μg DNA was achieved, which is 100-fold higher than the standard transformation procedure that yielded 5.3×103 transformants/μg DNA in the same strain.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    World journal of microbiology and biotechnology 15 (1999), S. 1-6 
    ISSN: 1573-0972
    Keywords: Bacteria ; conjugation ; DNA ; evolution ; gene transfer ; transduction ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The transfer of genetic information by transformation, conjugation and transduction in bacteria occurs frequently in nature. These diverse gene transfer mechanisms in bacteria are the result of evolution and are not linked to reproduction as in eukaryotic organisms. In this review, gene transfer in bacteria will be considered from an evolutionary perspective.
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  • 3
    ISSN: 1573-8221
    Keywords: human papilloma virus ; transformation ; actin ; fibronectin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Transfection of rat embryonic fibroblasts with E7 gene of type 16 human papilloma virus changed the cytoskeleton and cell-cell and cell-matrix interactions in two clones of transformed cells. Cell morphology and substrate-dependent proliferation were also changed.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Computer supported cooperative work 8 (1999), S. 63-93 
    ISSN: 1573-7551
    Keywords: activity theory ; action ; transformation ; expansive learning ; intervention ; visibilization ; health care ; medical records
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science
    Notes: Abstract Work is commonly made visible along two dimensions: the linear and the socio-spatial. Both are limited to depicting work in terms of relatively discrete actions. Activity theory introduces the crucial distinction between collective activity systems and individual actions. Expansive visibilization of collective activity systems offers a powerful intervention methodology for dealing with major transformations of work. The linear and the socio-spatial dimensions of work actions are seen in the broader perspective of a third, developmental dimension of work activity. Four steps are identified in a cycle of expansive visibilization, combining activity-level visions and action-level concretizations. The cycle is examined in detail as it unfolded in an intervention study at a children's hospital in Finland. It is concluded that expansive visibilization, driven by contradictions and seeking to reconceptualize the object and motive of work, is not a straightforward process which can be neatly controlled from above. Coherent analytical explanation and goal-setting may come only after the creation and practical implementation of innovative solutions.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    GeoJournal 49 (1999), S. 43-51 
    ISSN: 1572-9893
    Keywords: housing market ; suburbanisation ; transformation ; urban development ; urban renewal ; East Germany
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geography
    Notes: Abstract The aim of this paper is to analyse the main characteristics of post-socialist urban development in East Germany, especially the differences compared to urban development in other East and Central European countries. In spite of the many similar problems and processes in urban development, specific features of East Germany are characterised by the rapid growth of suburbia, especially in the first phase of transition, by the proceeding activities of urban renewal and revitalisation, and by a lower level of social polarisation and socio-spatial segregation as compared to other post-socialist countries. Important conditions for urban development in East Germany exist in special support programmes, high subsidies and other financial transfers as well as in engaged planning conceptions of the local authorities.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Cancer and metastasis reviews 18 (1999), S. 215-230 
    ISSN: 1573-7233
    Keywords: transformation ; tumour ; Frizzled ; Dishevelled ; glycogen synthase kinase-3β
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Wnt signalling is involved in a variety of mammalian developmental processes, including cell proliferation, differentiation and epithelial–mesenchymal interactions, through which they contribute to the development of tissues and organs such as the limbs, the brain, the reproductive tract and the kidney. Wnts are secreted ligands that control cell processes via at least two pathways, one of which, the ‘canonical’ Wnt signalling pathway, operates through the cytosolic stabilisation of a transcriptional co-factor, β-catenin. This is achieved by downregulating the activity of a β-catenin turnover complex. Evidence from tumour expression studies, transgenic animals and in vitro experiments suggests that inappropriate activation of the canonical Wnt signalling pathway is a major feature in human neoplasia and that oncogenic activation of this pathway can occur at many levels. Inappropriate expression of the Wnt ligand and Wnt binding proteins have been found in a variety of human tumours. Further downstream, dysregulation of the β-catenin turnover complex, by loss of the Adenomatous Polyposis Coli or Protein Phosphatase 2A proteins, or by activating mutations of β-catenin, has been found in several tumour types, and is believed to be a key step in neoplastic progression. Transcriptional targets of the Wnt pathway include the cellular oncogenes cyclin D1 and c-myc. Activation of the Wnt signalling pathway by various means can therefore be a primary cause in oncogenesis, affecting cell proliferation, morphology and contact inhibition, as well as co-operating with other oncogenes in multistep tumour progression.
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  • 7
    ISSN: 1573-6784
    Keywords: Bacillus ; plasmids ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A simple and easy method for the introduction of plasmid DNA into different species of Bacillus was developed. The method involves the suspension in a transformation buffer of nutrient agar grown cells in their late exponential phase and the addition of unpurified plasmid DNA. Transformants were obtained at a frequency of about 103 to 105 stable transformants per μg of plasmid DNA.
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  • 8
    ISSN: 1572-9729
    Keywords: abiotic ; biological ; cell-free extract ; chloroethane ; dechlorination ; 1,1-dichloroethane ; 1,1-dichloroethene ; digester ; methanogenic ; transformation ; 1,1,1-trichloroethane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Anaerobic transformations of 1,1,1-trichloroethane (TCA), 1,1-dichloroethane (DCA), and chloroethane (CA) were studied with sludge from a lab-scale, municipal wastewater sludge digester. TCA was biologically transformed to DCA and CA and further to ethane by reductive dechlorination. TCA was also converted to acetic acid and 1,1-dichloroethene (11DCE) by cell-free extract. 11DCE was further biologically converted to ethene. This pathway was confirmed by transformation tests of TCA, DCA and CA, by tests with cell-free extract, and by chloride release during TCA degradation. With cell-free extract, acetic acid accounted for approximately 90% of the TCA transformed; tests with live cells indicate that the fraction of TCA transformed by this pathway decreased with lower biomass. The dechlorination of DCA to CA and CA to ethane was not stoichiometric. A high rate of TCA removal was observed under the experimental conditions. The results indicate that removal of TCA in anaerobic digestion should be complete, but DCA and CA could persist in a normally operating digester.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology reporter 17 (1999), S. 323-331 
    ISSN: 1572-9818
    Keywords: Agrobacterium ; modular vector ; transformation ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Wheat (cv Chinese Spring) tissues were transformed using Agrobacterium tumefasciens and a new plasmid modular vector, pMVTBP. We constructed pMVTBP with unique restriction sites connecting (1) the CaMV 35S promoter, (2) a Kozak sequence, (3) the FLAG epitope, (4) the (His)6 epitope, (5) a coding region (for wheat TATA Binding Protein, wTBP) and (6) the CaMV 35S 3′UTR. This vector thus allows easy exchange of different regulatory or coding sequences. Explants of either germinating mature seeds, or immature embryos, were induced to callus for up to two weeks, treated with virulence-induced bacteria for one hour, then regenerated into plantlets. Transient expression of a GUS reporter gene, assayed at about one week, occurred in 10–12% of calluses. Expression of the FLAG-tagged wTBP was also detected, by immunostaining. Stable expression, by selective growth on geneticin, and by GUS expression at about six weeks, occurred in 1–2% of calluses, quite comparable to that achieved by other methods.
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  • 10
    ISSN: 1573-5079
    Keywords: Chlamydomonas reinhardtii ; LHC II phosphorylation ; mutagenesis ; Photosystem II redox control ; state 2 to state 1 transition ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Oxygenic photosynthetic organisms adapt to varying light conditions by changing the distribution of light energy between Photosystem II (PS II) and photosystem I (PS I) during so-called state transitions. To identify the genes involved in this process, we have exploited a simple chlorophyll fluorescence video-imaging technique to screen a library of nuclear mutants of Chlamydomonas reinhardtii for colonies grown on agar plates that are disturbed in their ability to regulate light energy distribution between PS I and PS II. Subsequent modulated fluorescence measurements at room temperature and 77 K fluorescence emission spectra confirmed that 5 mutants (0.025% of total number screened) were defective in state transitions. [32P]orthophosphate phosphorylation experiments in vivo revealed that in one of these mutants, designated stm1, the level of LHC II polypeptide phosphorylation was drastically reduced compared with wild type. Despite WT levels of PS I and PS II, stm1 grew photoautotrophically at reduced rates, compared with WT especially under low light conditions, which is consistent with an important physiological role for state transitions. Our results highlight the feasibility of video imaging in tandem with mutagenesis as a means of identifying the genes involved in controlling state transitions in eukaryotic photosynthetic organisms.
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  • 11
    ISSN: 1573-5028
    Keywords: constitutive expression ; GFP ; GUS ; Musa ; ScBV ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A 1369 bp DNA fragment (Sc) was isolated from a full-length clone of sugarcane bacilliform badnavirus (ScBV) and was shown to have promoter activity in transient expression assays using monocot (banana, maize, millet and sorghum) and dicot plant species (tobacco, sunflower, canola and Nicotiana benthamiana). This promoter was also tested for stable expression in transgenic banana and tobacco plants. These experiments showed that this promoter could drive high-level expression of the β-glucuronidase (GUS) reporter gene in most plant cells. The expression level was comparable to the maize ubiquitin promoter in standardised transient assays in maize. In transgenic banana plants the expression levels were variable for different transgenic lines but was generally comparable with the activities of both the maize ubiquitin promoter and the enhanced cauliflower mosaic virus (CaMV) 35S promoter. The Sc promoter appears to express in a near-constitutive manner in transgenic banana and tobacco plants. The promoter from sugarcane bacilliform virus represents a useful tool for the high-level expression of foreign genes in both monocot and dicot transgenic plants that could be used similarly to the CaMV 35S or maize polyubiquitin promoter.
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  • 12
    ISSN: 1573-5028
    Keywords: conditional lethal dominant gene ; Cre/loxP ; Nicotiana tabacum ; site-specific recombinase ; transformation ; transient expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic tobacco plants were produced that contained single-copy pART54 T-DNA, with a 35S-uidA gene linked to loxP-flanked kanamycin resistance (nptII) and cytosine deaminase (codA) genes. Retransformation of these plants with pCre1 (containing 35S transcribed cre recombinase and hygromycin (hpt) resistance genes) resulted in excision of the loxP-flanked genes from the genome. Phenotypes of progeny from selfed-retransformed plants confirmed nptII and codA excision and integration of the cre-linked hpt gene. To avoid integration of the hpt gene, and thereby generate plants totally free of marker genes, we attempted to transiently express the cre recombinase. Agrobacterium tumefaciens (pCre1) was cocultivated with leaf discs of two pART54-transformed lines and shoots were regenerated in the absence of hygromycin selection. Nineteen of 773 (0.25%) shoots showed tolerance to 5-fluorocytosine (5-fc) which is converted to the toxic 5-fluorouracil by cytosine deaminase. 5-fc tolerance in six shoots was found to be due to excision of the loxP-flanked region of the pART54 T-DNA. In four of these shoots excision could be attributed to cre expression from integrated pCre1 T-DNA, whereas in two shoots excision appeared to be a consequence of transient cre expression from pCre1 T-DNA molecules which had been transferred to the plant cells but not integrated into the genome. The absence of selectable marker genes was confirmed by the phenotype of the T1 progeny. Therefore, through transient cre expression, marker-free transgenic plants were produced without sexual crossing. This approach could be applicable to the elimination of marker genes from transgenic crops which must be vegetatively propagated to maintain their elite genotype.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 40 (1999), S. 711-717 
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; Arabidopsis thaliana ; binary vector ; T-DNA ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A streamlined mini binary vector was constructed that is less than 1/2 the size of the pBIN19 backbone (3.5 kb). This was accomplished by eliminating over 5 kb of non-T-DNA sequences from the pBIN19 vector. The vector still retains all the essential elements required for a binary vector. These include a RK2 replication origin, the nptIII gene conferring kanamycin resistance in bacteria, both the right and left T-DNA borders, and a multiple cloning site (MCS) in between the T-DNA borders to facilitate cloning. Due to the reduced size, more unique restriction sites are available in the MCS, thus allowing more versatile cloning. Since the traF region was not included, it is not possible to mobilize this binary vector into Agrobacterium by triparental mating. This problem can be easily resolved by direct transformation. The mini binary vector has been demonstrated to successfully transform Arabidopsis plants. Based on this mini binary vector, a series of binary vectors were constructed for plant transformation.
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  • 14
    ISSN: 1573-5044
    Keywords: ß-glucuronidase ; dendrobium ; hygromycin phosphotransferase ; orchid ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protocorms of orchid (Dendrobium hybrid) were transformed by microprojectile bombardment with a helium-pressured PDS 1000 particle gun. Gold particles coated with plasmid DNA containing ß-glucuronidase (GUS) and hygromycin phosphotransferase (Hpt) marker genes were used. Potentially transformed tissues were identified by active growth on MS medium supplemented with 50mg l-1 hygromycin. After 4–6 months of continuous selection, 15 hygromycin-resistant lines were recovered. Integration of transgenes into the genome of the transformed protocorms and plantlets were confirmed by GUS histochemical assay and Southern blot hybridization. The transgenic protocorms have gone through propagation for more than 8 months and maintained their transgenic characters. These results indicate that we have established a system for orchid transformation in a relatively high frequency and the transgenes are stably expressed in the transgenic plants.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 57 (1999), S. 207-210 
    ISSN: 1573-5044
    Keywords: biolistics ; gene expression ; haploid ; transformation ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using the PDS-1000/He Biolistic® Particle Delivery System, the microprojectile travel distance, rupture disk pressure and DNA/gold particle concentrations were assessed in order to optimise short and longer-term β-glucuronidase reporter gene expression in microspore-derived embryos of wheat. The effects were also evaluated of using sterile filter paper to support explants and treatment with a high osmoticum medium (0.2 M mannitol/0.2 M sorbitol or 0.4 M maltose). In the optimised procedure, wheat microspore-derived embryos (MDEs), were placed on filter paper and incubated on medium containing 0.4 M maltose, for 4 h pre- and 45 h post-bombardment. Five μl pAHC25 (0.75 mg ml-1 in TE buffer) was precipitated onto 25 μl gold particles (60 mg ml-1 in sterile water), using 20 μl spermidine (0.1 M) and 50 μl CaCl2 (2.5 M). The particles were centrifuged and resuspended in 75 μl absolute ethanol prior to the preparation of 6 macrocarriers. A microprojectile travel distance of 70 mm, a rupture pressure of 1300 p.s.i., and a vacuum of 29′′ Hg were employed. Maltose at 0.4 M in the support medium was the most important factor influencing GUS activity in bombarded tissues. GUS activity, 1 day post-bombardment, reached 52 ± 17 GUS-positive foci/MDE (mean ± s.e.m, n=3), with 17 ± 4 foci/MDE at 15 days, giving a 3.0-fold increase (p〈0.05) compared to expression in MDEs bombarded on medium without a high osmoticum treatment.
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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    Water, air & soil pollution 110 (1999), S. 57-66 
    ISSN: 1573-2932
    Keywords: DDT ; kinetic ; organic pollutant ; sediment ; sorption ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The overall objective of this study was to investigate the sorption kinetics of DDT in sediment under similar experimental conditions employed in corresponding toxicity studies for bentic organisms. A batch of aerated Schoonrewoerdse Wiel sediment, initially spiked with DDT, was sampled over a period of seven days. Concentrations of DDT, DDD and DDE were determined in both the solid and the solution phase in the sediment/water system after separation by centrifugation. It was found that the extractable amount of DDT decreased with increasing contact time. This can partly be explained in terms of transformation of DDT into DDD. Furthermore, the present applied extraction procedure seems to be less effective with increasing contact time, indicating an increase in binding strength of DDT with the sediment material. Finally, on the basis of DDT, DDE and DDD concentrations in both the solid phase and the solution phase, partition coefficients were calculated, which appeared to be independent of the contact time. This points at a very rapid equilibrating between DDT in pore water and in the extractable forms adsorbed at the solid phase.
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  • 17
    ISSN: 1573-2932
    Keywords: arid-zone soils ; field capacity ; fractionation ; heavy metals ; kinetics ; redistribution ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Solid-phase transformation of added Cd, Cu, Cr, Ni, Pb and Zn, in two arid-zone soils incubated in the field capacity moisture regime for one year, were studied. The heavy metals were fractionated into six empirically defined fractions using a selective sequential dissolution (SSD) protocol optimized for arid-zone soils. Each of these fractions was named based on the major soil component targeted for dissolution during the specific SSD step, but it is not assumed that they are mineralogically and chemically totally specific. The transformations of the metals in the two soils incubated at the field capacity regime were compared with those at the moisture saturation regime (Han and Banin, 1997). An initial fast stage of transformation of the soluble metals from the exchangeable (EXC) fraction to the less labile fractions (the carbonate (CARB) fraction for Cd, Pb, Zn, Ni and Cu, and the organic matter (OM) fraction for Cr, and to some extent Cu and Ni) occurred during the fractionation and within one hour after addition. This was followed by a second stage, involving long-term transformation processes of all metals: added Cd was transferred from the EXC into the CARB fraction; added Cr was transferred from the CARB to the OM fraction and Pb was transferred very slowly to the easily reducible oxide (ERO) fraction. Added Cu, Ni and Zn were transferred from the EXC and CARB fractions into the ERO fraction and to some extent OM and RO fractions. In Part I of this series, we reported that during incubation in the saturated moisture regime, Zn and Ni were transferred mainly into the RO and OM fractions. Cadmium, Cr and Pb underwent the same transformation pathways during the slow long-term process, with slightly different rates, in both water regimes. At low levels of addition, the incubated soils moved over one year towards a distribution similar to that of the native soil. At higher levels, the soils still remained removed from the quasi-equilibrium which characterized the native soil, even at the end of one year of incubation.
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  • 18
    ISSN: 1573-2932
    Keywords: atmospheric fate ; atmospheric transport ; deposition ; emission ; long-range transport ; pesticides ; registration ; remote area ; risk assessment ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The Health Council of the Netherlands organised an international workshop on the fate of pesticides in the atmosphere and possible approaches for their regulatory environmental risk assessment. Approximately forty experts discussed what is currently known about the atmospheric fate of pesticides and major gaps in our understanding were identified. They favoured a tiered approach for assessing the environmental risks of atmospheric dispersion of these chemicals. In the first tier a pesticide's potential for emission during application, as well as its volatilisation potential should be assessed. Estimates of the former should be based on the application method and the formulation, estimates of the latter on a compound's solubility in water, saturated vapour pressure and octanol/water partition coefficient. Where a pesticide's potential for becoming airborne exceeds critical values, it should be subjected to a more rigorous second tier evaluation which considers its toxicity to organisms in non-target areas. This evaluation can be achieved by calculating and comparing a predicted environmental concentration (PEC) and a predicted no-effect concentration (PNEC). By applying an extra uncertainty factor the PNEC can be provisionally derived from standard toxicity data that is already required for the registration of pesticides. Depending on the distance between the source and the reception area, the PEC can be estimated for remote areas using simple dispersion, trajectory type models and for nearby areas using common dispersion models and standard scenarios of pesticide use. A pesticide's atmospheric transport potential is based on factors such as its reaction rate with OH radicals. It should be used to discriminate between those compounds for which only the risks to nearby ecosystems have to be assessed, and those for which the risks to remote ecosystems also have to be determined. The participants were of the opinion that this approach is, in principle, scientifically feasible, although the remaining uncertainties are substantial. Further field and laboratory research is necessary to gain more reliable estimates of the physico-chemical properties of pesticides, to validate and improve environmental fate models and to validate the applicability of standard toxicity data. This will increase both the accuracy of and our confidence in the outcome of the risk assessment.
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  • 19
    Electronic Resource
    Electronic Resource
    Springer
    Journal of thermal analysis and calorimetry 56 (1999), S. 1133-1140 
    ISSN: 1572-8943
    Keywords: branched diamine ; melting ; polyamides ; polymorphism ; transformation ; WXRD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Our X-ray work of Dytek®-A, 2-methyl-pentamethylenediamine, containing polyamides shows polymorphism, whereas the polyamides with linear diamines do not. The polyamide of Dytek®-A and dodecanedioic acid, MPMD-12, is singled out for discussion and compared with the unbranched analogs of polyamides 6,12 and 5,12. Due to the presence of the -CH3 side group in the 2-position of the diamine, the polyamide MPMD-12 exhibits two stable crystal conformations. The new δ polymorph is not seen in linear polyamides 6,12 and 5,12. Studies by DSC polyamide MPMD-12 clearly illustrates at least two crystal forms, γ and δ, coexisting over a wide temperature range, and the isolation of each phase is possible by controlling temperature and time. The DMA modulus in the temperature region between the glass transition (or alpha relaxation) and melting transition shows strong dependence on the thermal history as demonstrated in a study of crystallization kinetics.
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  • 20
    Electronic Resource
    Electronic Resource
    Springer
    Journal of engineering mathematics 36 (1999), S. 241-254 
    ISSN: 1573-2703
    Keywords: flows in porous media ; transformation ; heat transfer ; drying bins ; conformal mapping.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mathematics , Technology
    Notes: Abstract The design of a drying or cooling store aims to provide an even airflow distribution, when aerated, for preservation purposes. The airflow in some curved bottom bins are studied in this paper. The flow is modelled, using Darcy's law. A generalized Schwarz-Christoffel transformation is employed to reduce the problem of computing streamlines and isobars of airflow to solving a single nonlinear equation for the flow angle along the wall. Corresponding to different bin shapes, a few computed streamlines and isobars of airflow are presented, showing the effect of changing bottom geometries on the air flow. Heat transfer in such bins is also investigated. Based on an analysis of the far field of airflow, finite-height bins are considered. Analytical solutions of the heat conduction equation in terms of streamlines and isobars are obtained.
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  • 21
    Electronic Resource
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    Springer
    Higher education 38 (1999), S. 275-290 
    ISSN: 1573-174X
    Keywords: academic staff ; curriculum change ; equity ; governance ; staff development ; student needs ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Nature of Science, Research, Systems of Higher Education, Museum Science
    Notes: Abstract In South Africa the restructuring of the higher education system and the transformation of higher education institutions are located within the country's broad political and socio-economic transition to democracy. This paper focuses particularly on institutional transformation, and pays attention to the implications of the process of transformation for academic staff. The following five interlinked and interdependent issues characterizing institutional transformation in South African higher education are identified: democratising the governance structures of institutions increasing access for educationally and financially disadvantaged students restructuring the curriculum focusing on developmental needs in research and community service redressing inequalities in terms of race and gender. Although the overall effect of institutional transformation is experienced rather negatively by many academic staff members, the paper concludes that academics have to be empowered by means of staff development to remain active partners in the transformation process.
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  • 22
    ISSN: 1572-9788
    Keywords: Triticum turgidum L. var. durum ; pasta wheat ; transformation ; seed protein modification ; flour quality improvement
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Particle bombardment has been used to transform three cultivars (L35, Ofanto, Svevo) and one breeding line (Latino × Lira) of durum wheat (Triticum turgidum L. var. durum). These varieties were co-transformed with plasmids containing selectable and scorable marker genes (bar and uidA) and plasmids containing one of two high-molecular-weight (HMW) glutenin subunit genes (encoding subunits 1Ax1 or 1Dx5). Ten independent transgenic lines were recovered from 1683 bombarded scutella (transformation efficiency thus 0.6%). Five lines expressed either subunit 1Dx5 or 1Ax1 at levels similar to those of endogenous subunits encoded on chromosome 1B. To identify the effects of the transgenes on the functional properties of grain, three lines showing segregation for transgene expression were used to isolate sibling T2 plants which were null or positive for the transgene product. Analysis of these plants using a small-scale mixograph showed that expression of the additional subunits resulted in increased dough strength and stability, demonstrating that transformation can be used to modify the quality of durum wheat for bread and pasta making.
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  • 23
    ISSN: 1572-9788
    Keywords: alternative oxidase ; antisense ; male-sterility ; tapetum-specific promoter ; tobacco ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The alternative oxidase of plant mitochondria is the terminal oxidase of the cyanide-insensitive respiratory pathway and is encoded by a nuclear gene. A 1 kb genomic fragment including exon 3 of the alternative oxidase was amplified by PCR from the genome of Arabidopsis thaliana. This fragment was connected to a tapetum-specific promoter in the antisense orientation and then introduced into tobacco. The pollen viability in three transgenic plants ranged from 2% to 60%. The reduced pollen viability cosegregated with the transgene in a selfed progeny. Immunolocalization of alternative oxidase protein in the immature flower bud section indicated that expression of alternative oxidase protein in tapetum of the transgenic plant was much lower than that of the non-transformant. The histological observation and protein gel-blot analysis showed that the development of pollen grains in the transgenic plant did not progress after the degradation of the tapetum, and the amount of alternative oxidase in pollen grains of the transgenic plant became lower than that of the non-transformant. These results suggested that the alternative oxidase activity in the tapetum has a significant effect on the pollen development.
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  • 24
    ISSN: 1572-9788
    Keywords: Cyamopsis tetragonoloba ; β-lactamase inhibitor ; sulbactam ; transformation ; transgene stability ; transgenic guar
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A procedure for transformation of the large-seeded endospermous legume guar (Cyamopsis tetragonoloba L.) and a study on transmission of the transgenes to offspring generations are presented. Using Agrobacterium tumefaciens with a T-DNA construct harbouring a β-glucuronidase gene (uidA) and a neomycin phosphotransferase gene (nptII), maximum transformation frequencies of cotyledonary explants were obtained using 145 mg/l kanamycin sulfate as selective agent. Carbenicillin and cefotaxime, used for the elimination of Agrobacterium after co-culture, displayed considerable toxicity to guar tissues but replacing most of these β-lactams by the non-phytotoxic β-lactamase inhibitor sulbactam as well as addition of thidiazuron and silver thiosulfate increased transformation frequencies up to 10-fold in total. The presence of the transgenes in the primary transformants was demonstrated by genomic DNA analysis of GUS-positive shoots. Chimaeric plants (5–10%) were identified by GUS analysis at the flowering stage and were discarded. Analysis of the R1 offspring from 17 independent transformants showed that in 41% of those, the uidA gene(s) was expressed and stably inherited consistent with Mendelian genetics. This was also found for the R2 and R3 generations of single copy transformants. On the other hand, a large proportion (47%) of the primary transformants gave R1 offspring in which 100% of the plants were GUS-negative. Analysis of these plants by PCR revealed that, at least, most of the transgene sequences were absent, suggesting that they had not been transmitted from the parent transformants. This occurred at similar high frequencies (40–50%) irrespective of the estimated copy number of the transgenes. Thus, major parts of the transgenes, even when present in multiple copies, displayed aberrant transmission, at a high frequency, in the process of going from the primary transformants to the first offspring generation.
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  • 25
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    European journal of plant pathology 105 (1999), S. 221-229 
    ISSN: 1573-8469
    Keywords: mutagenesis ; transformation ; plant disease ; recombination ; plant pathogenic fungi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Development of molecular techniques for phytopathogenic fungi aims at the identification of fungal genes whose products are essential for successful infection of the host plant. Initial approaches have relied on isolating candidate genes and generating null-mutations by homologous recombination. Unfortunately, the results of this strategy have not been overly successful. This has led to a search for alternatives which allow an unbiased identification of pathogenicity genes. One method, which has proved successful in several systems, is a tagging mutagenesis procedure termed restriction enzyme mediated integration (REMI). In this mini-review we describe this procedure and review its features and results of its use when applied to the identification of fungal genes required for disease development in planta.
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  • 26
    ISSN: 1573-9368
    Keywords: Agrobacterium tumefaciens ; ß‐glucuronidase ; lamiaceae ; lavandin ; neomycin phosphotransferase II ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Lavandin (Lavandula x Emeric ex Loiseleur) is an aromatic plant, the essential oil of which is widely used in the perfume, cosmetic, flavouring and pharmaceutical industries. The qualitative or quantitative modification of its terpenes‐containing essential oil by genetic engineering could have important scientific and commercial applications. In this study, we report the first Agrobacterium tumefaciens‐mediated gene transfer into lavandin. The transformation protocol was optimized by lengthening precultivation and cocultivation periods and by testing five different bacterial strains. We obtained transformed callus lines at a frequency of 40–70 with strains AGL1/GI, EHA105/GI and C58/GI. Transgenic shoots were regenerated from these kanamycin resistant calli and rooted on selective medium with 150 mg l-1 kanamycin. The final percentage of transgenic plants obtained varied from 3 to 9, according to the strain used, within 6 months of culture. The presence of the introduced β‐glucuronidase and neomycin phosphotransferase II genes was shown both by PCR and Southern blot analysis. Transgene expression was investigated using histoenzymatic β‐glucuronidase assays, leaf callus assays and RT‐PCR. Results showed that both β‐glucuronidase and neomycin phosphotransferase II genes were expressed at a high level in at least 41 of the transgenic plants regenerated. This efficient transformation strategy could be used to modify some genetic traits of lavandin (flower colour, pathogens resistance) and to study the biosynthesis of the major monoterpene components of its essential oil (linalool, linalyl acetate, camphor and 1,8‐cineole).
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  • 27
    ISSN: 1573-9368
    Keywords: Lactuca sativa ; Agrobacterium tumefaciens ; bialaphos ; phosphinothricin acetyltransferase ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Resistance to bialaphos, a broad-spectrum herbicide, was introduced into Lactuca sativa cv. Evola by Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strains 0310 and 1310, both carrying the bialaphos resistance (bar) and neomycin phosphotransferase (nptII) genes, were used for transformation. Primary transformants were selected on kanamycin sulphate-supplemented shoot regeneration medium. Integration of both transgenes was confirmed by non-radioactive Southern hybridisation. The hypervirulent plasmid ToK47 in A. tumefaciens strain 1310 generated multiple insertions of T-DNA in some transgenic plants; the absence of pToK47 (strain 0310) resulted in single gene inserts in all plants tested. Resistance to glufosinate ammonium was observed in axenic seedlings grown on medium supplemented with the herbicide at 5 mg l−1 and in glasshouse-grown plants sprayed with the compound at 300 mg l−1. Stable expression of the bar gene was observed in R2 generation plants. The kanamycin resistance of R1 seedlings was observed by germinating seeds on medium supplemented with 200 mg l−1 kanamycin sulphate. The presence of NPTII protein and PAT enzyme activity were demonstrated by ELISA and PAT enzyme assay respectively. Transgenes segregated in a Mendelian fashion in some plant lines in the R1 generation; herbicide resistance also segregated in the expected ratio in the R2 generation in most transgenic lines. This study confirmed that an agronomically important transgene can be integrated and stably expressed over several generations in lettuce.
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  • 28
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    GeoJournal 44 (1998), S. 215-224 
    ISSN: 1572-9893
    Keywords: cross-border region ; transformation ; regional economic development ; Poland ; Germany
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geography
    Notes: Abstract One of the major means to foster European integration is the establishment of border spanning regions (‘Euroregions’). This is particularly important on the Eastern borders of the EU, e.g. in Eastern Germany. There, however, a double transformation to post-socialist society is taking place, both inside and outside the EU. Tensions arise between objectives on local and higher political levels, intensified by totally different economic structures and access to EU funds on both sides of the border. This is particularly true for the case of the emerging Euroregion Viadrina. Problems in preserving old industrialised localities in East Germany (e.g. steel) and attempts to resurrect the urban fair place Frankfurt/Oder, clash with transition in agriculture and consumer industries and with new concepts in tourism development and environmental protection in the Polish border zone. In region building, political, economic and ideological goals compete with each other. Local initiatives and higher political governance may both support and hamper each other. The same holds true for the interdependence of cultural integration and economic development. The paper concludes that regional economic development can only be expected if, via the building of the Euroregion, the interplay of these factors leads to compromise and harmonization between the different parties involved.
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  • 29
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    GeoJournal 46 (1998), S. 263-269 
    ISSN: 1572-9893
    Keywords: Croatia ; decentralisation ; diversification ; industry ; innovation ; rural ; transformation ; urbanisation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geography
    Notes: Abstract Rural diversification in Croatia is well advanced because many rural families have been able to find work in secondary and tertiary activities without the need to migrate to the towns. Many rural settlements have now attained an urban character although there are regional variations, including a contrast between the continental zone with a relatively high level of commitment to agriculture and the coastal areas, with pronounced ‘deagrarisation’ where the ports and tourist resorts are well developed and the natural resource conditions for agriculture are poor. These variations are examined at the municipality level with reference to two key indicators: the share of nonagricultural population and the share of workers in the total active population. Four categories of socio-economic transformation are recognised: more urbanised, urbanised, less urbanised and rural. The main regional differences between the continental and coastal areas are confirmed with the latter showing a relatively high level of socio-economic transformation through the prominence of more highly urbanised municipalities.
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  • 30
    ISSN: 1572-9893
    Keywords: chamber of commerce ; economy ; experts ; institutions ; local government ; Romania ; rural ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geography
    Notes: Abstract This study evaluates some aspects of the socio-economic transformation of rural Romania with reference to the views of representative organisations (at national, regional and local levels) and other experts. Interviews conducted in ten communes of nine Romanian counties (‘judete’) focus attention on the advantages and disadvantages of system change experienced since 1989; the most important problems and constraints for future socio-economic change; and appropriate policies and perspectives for development in the immediate future. Wherever appropriate the claims of interviewees are substantiated through reference to statistics, drawn in many cases from Chambers of Commerce & Industry (CCI). Local level representatives presented much more negative views on recent change than their national and regional level counterparts, but all agreed on the crucial problem of capital shortage. Thus while specific programmes to assist rural areas are justified, they cannot fully succeed until the national economy is able to grow more rapidly and attract greater foreign investment.
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  • 31
    ISSN: 1572-9893
    Keywords: agriculture ; border region ; communities ; commuting ; conservation ; Romania ; tourism ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geography
    Notes: Abstract A most distinctive feature of the settlement pattern of the Brasov area is the extreme dispersal of mixed farming encountered in the western extreme of the county to the north and south of Zarnesti: the Bran and Poiana Marului areas. Here a system of peasant subsistence farming developed in a political borderland between the Habsburg and Ottoman Empires. Despite feudal pressures, the peasantry took all available opportunities to extend their independence including elaborate transhumance systems. And after seeing transfrontier commerce as a source of plunder, in the tradition of Balkan highway robbery within relatively unregulated spaces, the peasantry has profited through employment in factories, particularly during the communist period. However, the current recession in manufacturing is throwing the rural population back on limited land resources. Although farming assumes an important subsistence role which contributes to stability, the long-term survival of these communities will depend on new sources of income. Rural tourism has considerable potential and a promising start has been made in Bran. There are, however, constraints on the further development of the business and great attention will have to be given to the conservation of the environment in both the Bucegi Mountains and the Piatra Craiului where national park status is proposed.
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  • 32
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    Breast cancer research and treatment 47 (1998), S. 197-199 
    ISSN: 1573-7217
    Keywords: breast cancer ; insulin-like growth factor system ; IGF-I receptor ; IGF-II receptor ; binding proteins ; prognosis ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In 1992, a special issue of Breast Cancer Research and Treatment was devoted to the insulin-like growth factors and breast cancer. In that issue, identification of the key components of the IGF system was reviewed and their potential role in breast cancer growth was described. In this issue, we revisit the IGF system with particular attention to data that further supports their role in the growth regulation of breast cancer. Several new facets of the IGF system are described, and several laboratories have more clearly defined how each individual component of the IGF system may influence breast cancer biology.
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  • 33
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    Biotechnology techniques 12 (1998), S. 829-832 
    ISSN: 1573-6784
    Keywords: Pseudomonas oleovorans ; electroporation ; transformation ; poly(β-hydroxyalkanoate) ; alkane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract An electroporation procedure for the transformation of Pseudomonas oleovorans was developed using a model plasmid, pCN51. The optimal electrotransformation was achieved with cells harvested at 45 to 60 min of growth and concentrated to a cell density of 5 OD600nm, plasmid concentration of 6 μg per 100 μl of cell suspension, and a 0.1-cm gap-width cuvette. Electroporation was performed at the settings of 250 ω, 25μF and 2.5 kV. Transformation yields in the order of 103 colony-forming-unit per electroporation sample were obtained. This is a first report of the electroporation of the commercially valuable bacterium Ps. oleovorans. © Rapid Science Ltd. 1998
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  • 34
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    Plant molecular biology reporter 16 (1998), S. 129-131 
    ISSN: 1572-9818
    Keywords: Agrobacterium tumefaciens ; binary vector ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the construction of a binary vector for Agrobacterium tumefaciens-mediated transformation, pBIN20, which contains a superlinker region located between the left and right Ti border sequences. This vector, derived from pBI121, simplifies the cloning of plant expression cassettes and has been used in our laboratory to create lines of transgenic BY-2 tobacco cells. This new vector contains more than 20 unique restriction sites as well as the nptII selectable marker gene within the Ti-DNA borders.
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  • 35
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    Antonie van Leeuwenhoek 73 (1998), S. 147-153 
    ISSN: 1572-9699
    Keywords: transformation ; plasmid integration ; Phaffia rhodozyma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Stable red astaxanthin-producing transformants were obtained after genetic transformation of two Phaffia rhodozyma mutants. A yellow mutant, accumulating β-carotene, and an albino mutant, accumulating phytoene, from P. rhodozyma were transformed using a genomic library of wild-type strain UCD 67-385 in the pBluescript vector. Hybridization assays, using the pBluescript DNA as a radioactive probe, indicate integration of vector sequences into the genome of the transformants. Transformants DNA was digested with restriction endonucleases, ligated with T4 DNA ligase and then used to transform E. coli. Ampicillin resistant plasmids, containing 0.1, 0.2, and 2.5 kb DNA inserts of P. rhodozyma, were rescued from the yeast red transformants. The molecular analysis indicate that transformation has occurred by an integration event of donor DNA into the genome of the host strains.
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  • 36
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    Plant cell, tissue and organ culture 55 (1998), S. 175-181 
    ISSN: 1573-5044
    Keywords: Conifer ; transformation ; virulence genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract As a preliminary step in efforts to develop a successful protocol for Agrobacterium-mediated transformation of cotyledonary explants of Pinus pinea L. embryos, we tested the ability of embrionary exudates of this species to induce the expression of the virulence genes virA, virB, virC, virD, virE and virG in Agrobacterium tumefaciens containing vir: lacZ fusion constructs. The results obtained in the vir induction assay indicated the absence of bactericidal or bacteriostatic plant compounds affecting A. tumefaciens growth, and showed that cotyledonary and embrionary exudates of P. pinea are able to induce all virulence genes studied, except virG. The data suggest that A. tumefaciens can be used for gene transfer into this important forest and fruit species.
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  • 37
    ISSN: 1573-5028
    Keywords: plants ; positive selection ; selectable marker ; Thermoanaerobacterium thermosulfurogenes ; transformation ; xylose isomerase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The xylose isomerase gene (xylA) from Thermoanaerobacterium thermosulfurogenes (formerly Clostridium thermosulfurogenes) has been expressed in three plant species (potato, tobacco, and tomato) and transgenic plants have been selected on xylose-containing medium. The xylose isomerase gene was transferred to the target plant by Agrobacterium-mediated transformation. The xylose isomerase gene was expressed using the enhanced cauliflower mosaic virus (CaMV) 35S promoter and the Ω′ translation enhancer sequence from tobacco mosaic virus. Unoptimized selection studies showed that, in potato and tomato, the xylose isomerase selection was more efficient than the established kanamycin resistance selection, whereas in tobacco the opposite was observed. Efficiency may be increased by optimization. The xylose isomerase system enables the transgenic cells to utilize xylose as a carbohydrate source. It is an example of a positive selection system because transgenic cells proliferate while non-transgenic cells are starved but still survive. This contrasts to antibiotic or herbicide resistance where transgenic cells survive on a selective medium but non-transgenic cells are killed. The results give access to a new selection method which is devoid of the disadvantages of antibiotic or herbicide selection.
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  • 38
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    Plant molecular biology 38 (1998), S. 597-607 
    ISSN: 1573-5028
    Keywords: transgene silencing ; epigenetics ; transgene expression ; transgenic plants ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An irregular pattern of transgene silencing was revealed in expression and inheritance studies conducted over multiple generations following transgene introduction by microprojectile bombardment of allohexaploid cultivated oat (Avena sativa L.). Expression of two transgenes, bar and uidA, delivered on the same plasmid was investigated in 23 transgenic oat lines. Twenty-one transgenic lines, each derived from an independently selected transformed tissue culture, showed expression of both bar and uidA while two lines expressed only bar. The relationship of the transgenic phenotypes to the presence of the transgenes in the study was determined using (1) phenotypic scoring combined with Southern blot analyses of progeny, (2) coexpression of the two transgenic phenotypes since the two transgenes always cosegregated, and (3) reactivation of a transgenic phenotype in self-pollinated progenies of transgenic plants that did not exhibit a transgenic phenotype. Transgene silencing was observed in 19 of the 23 transgenic lines and resulted in distorted segregation of transgenic phenotypes in 10 lines. Silencing and inheritance distortions were irregular and unpredictable. They were often reversible in a subsequent generation of self-pollinated progeny and abnormally segregating progenies were as likely to trace back to parents that exhibited normal segregation in a previous generation as to parents showing segregation distortions. Possible causes of the irregular patterns of transgene silencing are discussed.
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  • 39
    ISSN: 1573-5028
    Keywords: Agrobacterium ; apple ; GFP ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To investigate early events of Agrobacterium-mediated transformation of apple cultivars, a synthetic green fluorescent protein gene (SGFP) was used as a highly sensitive, vital reporter gene. Leaf explants from four apple cultivars (‘Delicious’, ‘Golden Delicious’, ‘Royal Gala’ and ‘Greensleeves’) were infected with Agrobacterium EHA101 harboring plasmid pDM96.0501. Fluorescence microscopy indicated that SGFP expression was first detected 48 h after infection and quantitative analysis revealed a high T-DNA transfer rate. Plant cells with stably incorporated T-DNA exhibited cell division and developed transgenic calli, followed by formation of transgenic shoots at low frequencies. The detection of SGFP expression with an epifluorescence stereomicroscope confirmed the effectiveness of SGFP as a reporter gene for detection of very early transformation events and for screening of putative transformants. The efficiency of the transformation and regeneration process decreased ca. 10000-fold from Agrobacterium infection to transgenic shoot regeneration, suggesting that factors other than Agrobacterium interaction and T-DNA transfer are rate-limiting steps in Agrobacterium-mediated transformation of apple.
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  • 40
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; Avena sativa ; badnavirus promoter ; constitutive and vascular expression ; GUS staining ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Regions of the sugarcane bacilliform badnavirus genome were tested for promoter activity. The genomic region spanning nucleotides 5999–7420 was shown to possess promoter activity as exemplified by its ability to drive the expression of the coding region of the uidA gene of Escherichia coli, in both Avena sativa and Arabidopsis thaliana. In A. sativa, the promoter was active in all organs examined and, with the exception of the anthers where the expression was localized, this activity was constitutive. In A. thaliana, the promoter activity was constitutive in the rosette leaf, stem, stamen, and root and limited primarily to vascular tissue in the sepal and the silique. The transgene was inherited and active in progeny plants of both A. sativa and A. thaliana.
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  • 41
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    Environmental and ecological statistics 5 (1998), S. 197-222 
    ISSN: 1573-3009
    Keywords: kriging ; non-separable space-time correlation ; spatial scale ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract We present an approach to estimate hourly grid-cell surface ozone concentrations based on observations from point monitoring sites in space, for comparison with grid-based results from the SARMAP photochemical air-quality model for a region of northern California. Statistical estimation is carried out on a transformed (square root) scale, followed by back-transforming to the original scale of ozone in parts per billion, adjusting for bias and variance. We estimate a spatially-varying diurnal mean structure and a non-separable space-time correlation structure on the transformed scale. Temporal pre-whitening is followed by modelling of a spatially non-stationary, diurnally-varying spatial correlation structure using a spatial deformation approach. Comparisons of SARMAP model results with the estimated grid-cell ozone levels are presented.
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  • 42
    ISSN: 1572-9788
    Keywords: Bt genes ; transformation ; protection against insects ; cry1Ia5
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A cry1Ia5 insecticidal toxin coding gene has been cloned from an Indian isolate of Bacillus thuringiensis. Sequence analyses of the cry1Ia5 gene revealed the absence of potential polyadenylation signal sequences thus making it a suitable candidate for expression in plants without extensive modification. This possibility was examined by subcloning the cry1Ia5 gene into a plant expression vector and then transferring it to Nicotiana tabacum through Agrobacterium-mediated transformation. Our results demonstrate that N. tabacum with a stably integrated native cry1Ia5 gene afforded complete protection against predation by Heliothis armigera. Forty three percent of the transgenic plants displayed a high level of protection against insect predation. The protection obtained in transgenic plants with the cry1Ia5 gene was comparable to that obtained with the synthetically modified cry1A(b) or cry1A(c) genes. The results demonstrate that novel insecticidal genes already exist in nature that do not require extensive modifications for efficient expression in plants.
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  • 43
    ISSN: 1572-9788
    Keywords: Beta vulgaris ; mannose selection ; phosphomannose isomerase ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Various factors affecting mannose selection for the production of transgenic plants were studied using Agrobacterium tumefaciens-mediated transformation of sugar beet (Beta vulgaris L.) cotyledonary explants. The selection system is based on the Escherichia coli phosphomannose isomerase (PMI) gene as selectable gene and mannose as selective agent. Transformation frequencies were about 10-fold higher than for kanamycin selection but were only obtained at low selection pressures (1.0–1.5 g/l mannose) where 20–30% of the explants produced shoots. The non-transgenic shoots were eliminated during the selection procedure by a stepwise increase in the mannose concentration up to 10 g/l. Analysis of the transformed shoots showed that the PMI activity varied from 2.4 mU/mg to 350 mU/mg but the expression level was independent of the selection pressure. Complete resistance to mannose of transformed shoots was observed already at low PMI activities (7.5 mU/mg). Genomic DNA blot analysis confirmed the presence of the PMI gene in all transformants analysed. The possible mode of action of mannose selection compared to other selection methods is discussed.
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  • 44
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    Molecular breeding 4 (1998), S. 531-541 
    ISSN: 1572-9788
    Keywords: Agrobacterium ; Brassica oleracea ; cauliflower ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We have developed an efficient and simpler method for genetic transformation and regeneration of cauliflower, Brassica oleracea var. botrytis plants. Explants from 4-day old seedlings were inoculated and cocultivated with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector with the neomycin phosphotransferase-II gene under the regulatory control of nopaline synthase promoter and terminator sequences, permitting transformed shoots to be selected on kanamycin containing medium. After three months rooted transformed plantlets were successfully transferred and grown under glasshouse conditions. Higher numbers of transformed plants were obtained from cotyledon than hypocotyl explants, presumably indicating cotyledons of cauliflower are more amenable to genetic transformation. Integration and expression of the introduced transgene were analysed by DNA gel blot and PCR analysis and NPT-II expression assay. Factors influencing transformation efficiency include explant age, concentration of bacterium used for infection, duration of infection and cocultivation with Agrobacterium. Transgenic plants of three commercial genotypes of cauliflower were produced using this method. We also show that introduction of antisense Bcp1 (pollen-specific gene) linked to a pollen-specific promoter (Lat52) resulted in the expected sterility of 50% pollen carrying this transgenic construct.
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  • 45
    ISSN: 1572-9788
    Keywords: glycerol-3-phosphate acyltransferase ; phosphatidylglycerol ; chilling tolerance ; transformation ; fatty acid composition ; Oryza sativa L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The chilling sensitivity of several plant species is closely correlated with the levels of unsaturation of fatty acids in the phosphatidylglycerol (PG) of chloroplast membranes. Plants with a high proportion of unsaturated fatty acids, such as Arabidopsis thaliana, are resistant to chilling, whereas species like squash with only a low proportion are rather sensitive to chilling. The glycerol-3-phosphate O-acyltransferase (GPAT) enzyme of chloroplasts plays an important role in determining the levels of PG fatty acid desaturation. A cDNA for oleate-selective GPAT of Arabidopsis under the control of a maize Ubiquitin promoter was introduced into rice (Oryza sativa L.) using the Agrobacterium-mediated gene transfer method. The levels of unsaturated fatty acids in the phosphatidylglycerol of transformed rice leaves were found to be 28% higher than that of untransformed controls. The net photosynthetic rate of leaves of transformed rice plants was 20% higher than that of the wild type at 17°C. Thus, introduction of cDNA for the Arabidopsis GPAT causes greater unsaturation of fatty acids and confers chilling tolerance of photosynthesis on rice.
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  • 46
    ISSN: 1572-9788
    Keywords: Indica rice ; cell suspension ; transformation ; Xa21 ; bacterial leaf blight
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The agronomically important Indica (group 1) rice varieties IR64, IR72, hybrid restorer line Minghui 63, and BG90-2 were co-transformed by microbombardment of embryogenic suspensions with plasmids that contain the Xa21 gene which confers resistance to Xanthomonas oryzae pv. oryzae and the hph gene for resistance to hygromycin B. Six of the 55 transgenic R0 plant lines containing the Xa21 gene displayed high levels of resistance to the pathogen, and no partial resistance was observed. The trait was stably inherited in subsequent generations, and transgenic plants are currently in field tests. The ability to transfer agronomically important genes into elite Indica rice varieties demonstrates the applicability of genetic engineering for the agronomic improvement of rice.
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  • 47
    ISSN: 1572-9788
    Keywords: antisense ; chalcone synthase ; flower colour ; lisianthus ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Three cultivars of lisianthus (Eustoma grandiflorum (Grise.)) were transformed with a homologous antisense CHS cDNA via Agrobacterium-mediated transformation. Over 50% of the transgenics derived from the purple flowering lines exhibited an altered flower colour pattern ranging from small streaks of white on the wild-type purple background through to completely white flowers. A significant portion of the transgenic lines showed unstable phenotypes. Northern and biochemical analysis showed that the altered flower patterns were associated with a loss of CHS gene transcript and a corresponding loss of CHS enzyme activity. In the white flowering line the level of total flavonoids was reduced to ca. 2.0% of the wild-type level. Some of the transgenic plants also exhibited alterations in flower form such as the formation of frilled petal tips and reduced flower opening. Several of the new patterned lines are being evaluated for stability and possible commercial release.
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  • 48
    ISSN: 1572-9788
    Keywords: amylose ; antisense RNA ; endogenous allele ; Solanum tuberosum ; T-DNA insertion ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The T-DNA composition was analysed of twelve potato genotypes obtained after transforming a tetraploid cultivar with an antisense granule-bound starch synthase (GBSSI) gene. In five transformants (labelled TB50 nos.) the antisense GBSSI gene was driven by the CaMV 35S promoter, while in the remaining seven (labelled TBK50 nos.) the GBSSI promoter was used. In these twelve transformants the antisense effect on amylose production in potato tuber starch ranged from complete suppression to no discernible inhibition, and the number of T-DNA insertions ranged from one to at least fifteen. The antisense effect of individual T-DNA loci in progeny of these transformants was studied. Progeny containing a single T-DNA showed no inhibition of GBSSI activity. Only multiple, linked T-DNA insertions resulted in substantial antisense inhibition. T-DNA fragments present in duplex in selfed progeny resulted in a larger antisense effect than that in the parent (which contained the T-DNA insertions in simplex). Furthermore, the antisense effects of some T-DNA-containing linkage groups were influenced by the composition of endogenous GBSSI alleles. For practical breeding this implies that (1) the efficiency of obtaining primary potato transformants showing complete inhibition of GBSSI gene expression by antisense RNA is genotype-dependent, and (2) many transformants have to be produced per genotype to be able to select plants with maximum suppression of GBSSI and a minimum number of T-DNA loci.
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  • 49
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    Social indicators research 43 (1998), S. 197-209 
    ISSN: 1573-0921
    Keywords: transformation ; anomie ; social integration ; state ; quality of life
    Source: Springer Online Journal Archives 1860-2000
    Topics: Sociology
    Notes: Abstract The weakening of social integration and anomie are unavoidable in the transformation of societies. The effect is a decrease of quality of life accompanied by disenchantments, aggressiveness and escapism. In some countries in Eastern Europe like Bulgaria the anomie effects of transformation became particularly strong. The major reason is the political instability. The dissolution of the previous state-centered over-integration of society developed into a dissolution of major mechanisms of political integration. The prospects for improvement of quality of life are focused on the balance of economic, political and cultural re-integration of Bulgarian society.
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  • 50
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    Euphytica 100 (1998), S. 219-223 
    ISSN: 1573-5060
    Keywords: cereals ; wheat ; transformation ; genetic modification ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A method for efficient genetic transformation of wheat has been developed using immature embryos as targets for microprojectile-mediated gene transfer and a helium driven particle delivery system. Screening and selection of transgenic cells, somatic embryos and regenerated plants are performed with the gus-gene and the phosphinothricin acetyl transferase (bar) gene coding for Basta-resistance as the selectable marker. On average, one fertile transgenic plant can be obtained from about 100 microprojectile treated, immature embryos. The number of integrated copies of the transferred gene ranges from 1 up to about 10. Stable integrated genes are inherited in most of the transgenic lines in a normal mendelian fashion segregating 3:1 in the F2. Homozygous, as well as heterozygous, lines have been followed and analysed genetically at the molecular level and up to F5. Apart from normal stable gene expression, examples have also been found which showed a loss of gene activity or unexpected segregation pattern. For applied aspects, different genes are transferred aiming for improved disease resistance, modification of quality, or other characteristics. First results from these transgenic lines are reported, and problems still existing with the production of stable transgenic wheat lines are discussed.
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  • 51
    ISSN: 1573-9368
    Keywords: chimaera ; iterative culture ; regeneration ; strawberry ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic plants of strawberry cultivar Totem were developed by Agrobacterium-mediated transformation using a plasmid vector containing gus and nptII genes. Parallel experiments were carried out with and without repeated subculturing (iterative cultures) for generation of transgenic shoots on selection medium. The selection levels in the non-iterative pathway were kept constant, while in the iterative protocol, stepwise increase of selection pressure was applied at different stages of tissue growth. Rooted transgenic plants obtained via both protocols were outplanted in soil. Random leaf samples of greenhouse-grown transgenics were analysed for the presence of gus gene sequences by Southern hybridization as well as gus expression on leaf and petiole tissues by X-Gluc histological assay. Random leaf samples analysed from individual transgenic events developed under iterative culture were positive for the gus insert as verified by Southern analysis confirming the presence of transgenes and lack of chimaeras. Leaf samples of the transgenic events from the non-iterative protocol were either positive or negative on Southern analysis indicating the chimaeric nature of the transgenic plants. The absence of gus sequences in the transgenic plants grown under the non-iterative protocol reinforced the necessity of iterative cultures along with stepwise increase in selection levels for generating non-chimaeric transgenics in strawberry. The gus expression was highly variable, irrespective of the iterative or non-iterative protocol used for transformation. We conclude that strawberry is highly prone to develop chimaeric transgenics if derived from primary regenerants and that the iterative culture technique effectively converts chimaeras to pure line transgenic plants
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  • 52
    ISSN: 1573-9368
    Keywords: sweet orange ; Citrus ; woody ; transformation ; Agrobacterium ; mature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Regeneration and transformation systems from mature plant material of woody fruit species have to be achieved as a necessary requirement for the introduction of useful genes into specific cultivars and the rapid evaluation of resulting horticultural traits. We report here, for the first time, a procedure for genetic transformation and regeneration of mature tissues of woody plants that overcomes the long juvenile periods and high heterozygosity that are characteristic of most of these species. An improved regeneration frequency from mature explants was obtained by invigoration of the plant material through grafting of mature buds on juvenile seedlings. Co-cultivation of the explants in feederplates after inoculation with Agrobacterium tumefaciens resulted in enhanced transformation frequencies. Furthermore, in vitro shoot-tip grafting of the regenerated mature shoots on seedling rootstocks provided a rapid and efficient system for plant production. Citrus is the most extensivel y grown fruit crop worldwide and sweet orange (Citrus sinensis L. Osbeck) accounts for approximately 70% of the Citrus total production. Mature transgenic sweet orange plants have been obtained, which flowered and bore fruit in 14 months
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  • 53
    ISSN: 1573-9368
    Keywords: Saccharum ; Agrobacterium ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This is the first successful report of the recovery of morphologically normal transgenic sugarcane plants from co-cultivation of calluses with Agrobacterium tumefaciens. Transformation frequencies (total of transgenic plants/number of cell clusters) were between 9.4 × 10−3 and 1.15 × 10−2. In our experiments, both LBA4404 (pTOK233) and EHA101 (pMTCA3IG), carrying a super-binary vector or supervirulent strain, respectively, were successful for sugarcane transformation. We found that three main factors: (1) the use of young regenerable calluses as target explants; (2) induction and/or improvement of the A. tumefaciens virulence system with sugarcane cell cultures and (3) pre-induction of organogenesis or somatic-embryogenesis-like sexual embryos, seem to be crucial in order to increase the cells competence for T-DNA transfer process. Patterns generated by Southern hybridization confirmed that T-DNAs were randomly integrated into sugarcane genome without th e persistence of A. tumefaciens in the transgenic plants
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  • 54
    ISSN: 1573-9368
    Keywords: Oryza sativa ; particle bombardment ; transformation ; transgenic rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We developed a practical and efficient gene transfer system for indica rice utilizing mature-seed derived explants and a simple bombardment device which uses compressed helium for accelerating DNA-coated metal particles. Unlike instruments which have been described in the literature previously, this new bombardment device, which is an improvement of the particle inflow concept, does not require vacuum. This attribute simplifies the transformation procedure significantly and it makes rice transformation technology accessible to laboratories which may not have the resources to invest in more expensive particle bombardment instruments. We determined experimentally that we could recover transgenic rice plants utilizing three different particle bombardment instruments at comparable frequencies.
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  • 55
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    Bognor Regis [u.a.] : Wiley-Blackwell
    Journal of Polymer Science Part A: Polymer Chemistry 36 (1998), S. 2209-2214 
    ISSN: 0887-624X
    Keywords: cationic polymerization ; anionic polymerization ; transformation ; samarium ; block copolymerization ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Transformation of the cationic growing center of living poly(tetrahydrofuran) [poly(THF)] into an anionic one was achieved in high efficiency (62%) by the end-capping of living poly(THF) with potassium iodide followed by the reduction with bis(pentamethylcyclopentadienyl)samarium (Cp*2Sm), whereas the direct reduction with Cp*2Sm without the end-capping resulted in the formation of poly(THF) with pentamethylcyclopentadienyl group at the terminal. The increase in the molecular weight of poly(THF) after the reduction was observed, which indicates the presence of the dimerization of poly(THF) during the reduction. The polymerization of a variety of electrophilic monomers including δ-valerolactone, 2-oxo-1,3-dioxane, and alkyl methacrylates with the macroanion provided good yields of the corresponding block copolymers consisting of both cationically and anionically polymerizable monomers. © 1998 John Wiley & Sons, Inc. J. Polym. Sci. A Polym. Chem. 36: 2209-2214, 1998
    Additional Material: 4 Ill.
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  • 56
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    Yeast 14 (1998), S. 565-571 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; electroporation ; transformation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Pretreatment of yeast cells with lithium acetate (LiAc) and dithiothreitol (DTT) enhances the frequency of transformation by electroporation. The method shows improvements of 6-67-fold in wild-type strains derived from commonly used Saccharomyces cerevisiae genetic backgrounds. In addition, 15-300-fold improvement in transformation frequency was achieved with several mutant strains of S. cerevisiae that transformed poorly by conventional procedures. Both DTT and lithium acetate were necessary for maximal transformation frequencies. Pretreatment with lithium and DTT also resulted in an ∼3·5-fold increase in the electroporation transformation frequency of the pathogenic fungus Candida albicans. © 1998 John Wiley & Sons, Ltd.
    Additional Material: 1 Ill.
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  • 57
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    Yeast 14 (1998), S. 1017-1025 
    ISSN: 0749-503X
    Keywords: Arxula adeninivorans ; AILV1 ; threonine deaminase ; transformation ; homologous integration ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ILV1 gene of the yeast Arxula adeninivorans LS3 (AILV1) has been cloned from a genomic library, characterized and used as an auxotrophic selection marker for transformation of plasmids into this yeast. One copy of the gene is present in the Arxula genome, comprising 1653 bp and encoding 550 amino acids of the threonine deaminase. The protein sequence is similar (60·55%) to that of the threonine deaminase from Saccharomyces cerevisiae encoded by the gene ILV1. The protein is enzymatically active during the whole period of cultivation, up to 70 h. Maximal activities, as well as protein concentrations of this enzyme, were achieved after cultivation times of 20-36 h.The AILV1 gene is a suitable auxotrophic selection marker in transformation experiments using an Arxula adeninivorans ilv1 mutant and a plasmid containing this gene, which is fused into the 25S rDNA of Arxula adeninivorans. One to three copies of the linearized plasmid were integrated into the 25S rDNA by homologous recombination. Transformants resulting from complementation of the ilv1 mutation can be easily and reproducibly selected and in addition are mitotically stable. Therefore, the described system is preferred to the conventional selection for hygromycin B resistance. © 1998 John Wiley & Sons, Ltd.
    Additional Material: 5 Ill.
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  • 58
    ISSN: 1432-0789
    Keywords: Cyanobacteria ; Soil inoculation N ; transformation ; Inorganic N ; Easily oxidizable N ; Hydrolysable N ; Non-hydrolysable N ; Wetland rice Farmyard manure ; Oryza sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A field experiment was conducted with wetland rice (Oryza sativa cv. IR-36) in a sandy clay loam soil (Entisol) to study the effect of inoculation with a soil-based mixed culture of four diazotrophic cyanobacteria,Aulosira fertilissima, Nostoc muscorum, N. commune andAnabaena spp., on the N-flux in inorganic NH4 ++NO3 −+ NO2 −), easily oxidizable, hydrolysable and non-hydrolysable forms of N in soil during vegetative growth periods of the crop. Effects on grain and straw yield and N uptake by the crop were estimated. The effects of applying urea N and N as organic sources, viz.Sesbania aculeata, Neem (Azardirachta indica) cake and FYM, each at the rate of 40 kg N ha−1, to the soil were also evaluated. Inoculation significantly increased the release of inorganic N, evidenced by its increased concentrations either in soil or in soil solution. However, such increases rarely exceeded even 4% of total N gained in different froms in the soil system by inoculation during the vegetative growth stages of the rice plant, when the nutritional requirement of the plants is at a maximum. Most of the N2 fixed by cyanobacteria remained in the soil as the hydrolysable form (about 85%) during this period. Inoculation caused an insignificant increase in grain (8%) and straw (11%) yield, which was, however, accompanied by a significant increase in N uptake by the grain (30%) and an increase in total uptake of 15.3 kg N ha 1. Such beneficial effects of inoculation varied in magnitude with the application of organic sources, with farmyard manure (FYM) being the most effective. Application of urea N, on the other hand, markedly reduced such an effect.
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  • 59
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    World journal of microbiology and biotechnology 14 (1997), S. 7-10 
    ISSN: 1573-0972
    Keywords: Biodegradation ; morpholine ; plasmid ; Pseudomonas fluorescens ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A fast-growing Pseudomonas fluorescens CAS102, isolated by enrichment technique from polluted soil, effectively utilized morpholine as the energy source. The strain was able to grow in high concentrations of morpholine but accumulation of ammonia inhibited its growth and complete mineralization. The molar conversion ratio of morpholine to ammonia was 1:0.82. The organism harboured a single, multiple antibiotic- and heavy metal-resistance 140kb plasmid which was resistant to curing. Transformation studies showed that the morpholine degradative phenotype was expressed only in Pseudomonas putida and not in Escherichia coli. Growth studies on different degradative intermediates of morpholine suggested that plasmid-encoded genes were involved in the heterocyclic ring cleavage and the remaining reactions were mediated by chromosomal genes.
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  • 60
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    World journal of microbiology and biotechnology 14 (1997), S. 271-276 
    ISSN: 1573-0972
    Keywords: Biodegradation ; 3-chlorobenzoic acid ; degradative plasmid ; Pseudomonas putida ; rec− Escherichia coli DH5 ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 61
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    Biologia plantarum 40 (1997), S. 507-514 
    ISSN: 1573-8264
    Keywords: herbicide resistance ; PCR ; Phaseolus vulgaris ; regeneration ; Southern blotting ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We obtained transformed bean shoots by electroporation of intact bean cells with the plasmid pDPG165 containing bar gene conferring herbicide resistance to plants. Transformed shoots were selected from electroporated callus on herbicide containing media. Data of molecular analysis (PCR and Southern blotting) confirmed the insertion of bar gene in the genome of herbicide resistant shoots. Detailed procedures for obtaining regenerative bean callus, optimization of electroporation of intact cells and transgenic shoots are given.
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  • 62
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    Pharmaceutical research 14 (1997), S. 750-756 
    ISSN: 1573-904X
    Keywords: DNA delivery ; cells ; sonication ; cavitation ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. Ultrasonic methods have considerable potential for the introduction of macromolecules into cells. In this paper we demonstrate that, under controlled conditions, application of 20kHz ultrasound to a suspension of yeast cells facilitates the delivery of plasmid DNA into these cells. Methods. Aliquots of growing yeast cells (Saccharomyces cerevisae, strain AH22) were suspended in buffer and exposed to 20kHz ultrasound from a laboratory (probe-type) sonicator in the presence of microgram quantities of plasmid DNA. Efficiency of DNA delivery was scored as the number of cells transformed. Results. Cell transformation was optimal at 30 seconds sonication using an output of 2.0 watts and resulted in a 20 fold enhancement over control values. At extended sonication times, fewer cells showed evidence of transformation because of reduced cell viability. The increased DNA uptake and the decreased cell viability were both attributable to acoustic cavitation events during sonication. The extent of acoustic cavitation was measured and it was found that there was an increase in cavitation events with increased sonication time. Cell viability was shown to be directly related to the number of cavitation events. The effects of sonication on plasmid DNA were investigated and indicated that the structural integrity of plasmid DNA was unaffected by the sonication conditions employed. Conclusions. Under controlled conditions, ultrasound is an effective means of delivering plasmid DNA into cells. The subsequent expression of DNA molecules in cells depends upon a balance between transient cell damage and cell death.
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  • 63
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    Catalysis letters 49 (1997), S. 121-128 
    ISSN: 1572-879X
    Keywords: cresols ; transformation ; isomerization ; disproportionation ; zeolites HY
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The transformation of m- and o-cresol has been investigated on a series of HY zeolites with Si/Al ratio in the range of 4.5 to 55, at 380°C and atmospheric pressure. It was found that the zeolite activity increased but stability decreased with NAl for both m- and o-cresol. For Si/Al = 4.5 the zeolite activity was similar for both reactants, but as Si/Al ratio increased, o-cresol became relatively more reactive than m-cresol, and for all Si/Al ratios the catalysts were more stable in the m-cresol transformation. Both m- and o-cresol were converted mainly through two parallel reactions - isomerization and disproportionation. The disproportionation reaction was predominant, and more important in the case of o-cresol. The isomerization/disproportionation selectivity increased for m-cresol and decreased for o-cresol with NAl. Catalyst deactivation favoured disproportionation in the case of o-cresol and isomerization in the case of m-cresol, except for o-cresol conversion on HY(4.5), where both reactions were equally affected. Isomerization selectivity for m-cresol (p/o) was found to be higher than the equilibrium value and to increase with catalyst deactivation; while for o-cresol, the isomerization ratio (m/p) did not change with time on stream for HY(4.5) and HY(16.6) and decreased for HY(55). Disproportionation selectivity (ph/∑x) was equal to unity independently of Si/Al ratio or catalyst deactivation.
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  • 64
    ISSN: 1572-994X
    Keywords: adenovirus type 12 ; E1A ; transformation ; p300 binding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The early region 1A (E1A) gene of highly oncogenic adenovirus type 12 (Ad12) was analyzed for transforming activity and protein binding using specific mutations. The Ad12 E1A proteins were found to bind p300 protein mainly within the CR1 region, although mutations that affect both p300 binding and transformation were identified in both the CR1 and the N-terminal region. The most critical mutation dlf89 located in the CR1 region was further dissected by point mutations and the results identified 68S as the most critical for transformation and 67E as the most critical for p300 binding. Specific mutations that retain p300 binding but impair transcriptional repression of a viral enhancer were also identified in both the N-terminal and CR1 regions.
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  • 65
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    International journal of value-based management 10 (1997), S. 9-29 
    ISSN: 1572-8528
    Keywords: business ethics ; Jewish ; religious ; community ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Economics
    Notes: Abstract This paper examines the method of Jewish business ethics. MichaelWalzer, in his work, Interpretation and Social Criticism (1987), suggeststhree common and important approaches to moral philosophy. He labels thesethe path of discovery, the path of invention, and the path ofinterpretation. The first part of this paper argues that Jewish businessethics is best thought of in terms of interpretation. Without question, thereligious ethicist immediately recognizes Walzer‘s metaphor of the moralworld as a ’home occupied by a single family over many generations... ‘as his own. Ethical arguments from a Jewish perspective must of necessityhave a ’lived-in quality‘ and always make reference to and are based on the’memory-laden objects and artifacts.‘ The second part of the paper exploressome of the implications of Jewish business ethics as interpretation.
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  • 66
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    Euphytica 94 (1997), S. 83-91 
    ISSN: 1573-5060
    Keywords: Agrobacterium tumefaciens ; Beta vulgaris ; 5-enolpyruvylshikimate-3-synthase ; glyphosate ; herbicide ; sugar beet ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Sugar beet (Beta vulgaris L.) lines transformed with the 5-enolpyruvylshikimate-3-phosphate synthase gene (CP4 EPSPS) from Agrobacterium sp. CP4 and a glyphosate oxidase reductase gene (GOX) also isolated from bacteria resulted in the development of lines highly tolerant to glyphosate. Glyphosate (N-phosphonomethyl-glycine) is the active ingredient in Roundup®, herbicide. The EPSPS enzyme is involved in the biosynthesis of aromatic amino acids. Glyphosate binds irreversible to the EPSPS and inhibits the pathway. GOX degrades glyphosate into non-toxic compounds. 260 independent transformants have been evaluated in greenhouse and field trials for tolerance to Roundup® in 1993 and 1994. Variation of tolerance was recorded between different transformants, ranging from complete susceptibility to full tolerance. The Agrobacterium tumefaciens mediated transformation resulted in a negative correlation between copy number of the T-DNA insert and the level of tolerance to the herbicide. Transformants which contain a single copy insert showed tolerance to higher doses of glyphosate than transformants with multiple copies. Two transgenic lines were identified that showed agronomically useful tolerance to glyphosate.
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  • 67
    ISSN: 1573-5079
    Keywords: Chlamydomonas reinhardtii ; mutagenesis ; photoinhibition ; Photosystem II ; repair cycle ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The work outlines the isolation of transformant Chlamydomonas reinhardtii cells that appear to be unable to repair Photosystem II from photoinhibitory damage. A physiological and biochemical characterization of three mutants is presented. The results show differential stability for the D1 reaction center protein in the three mutants compared to the wild type and suggest lesions that affect different aspects of the Photosystem II repair mechanism. In the ag16.2 mutant, significantly greater amounts of D1 accumulate in the thylakoid membrane than in the wild type under steady-state growth conditions, and D1 loss is significantly retarded in the presence of the protein biosynthesis inhibitor chloramphenicol. Moreover, aberrant electrophoretic mobility of D1 in the ag16.2 suggests that this protein is modified to an as yet unknown configuration. These results indicate that the biosynthesis and/or degradation of D1 is altered in this strain. A different type of mutation occurred in the kn66.7 and kn27.4 mutants of C. reinhardtii. The stability of D1 declined much faster as a function of light intensity in these mutants than in the wild type. Thereby, the threshold of photoinhibition in these mutants was significantly lower than that in the wild type. It appears that kn66.7 and kn27.4 are similar conditional mutants, with the only difference between them being the amplitude of the chloroplast response to the mutation and the differential sensitivity they display to the level of irradiance.
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  • 68
    ISSN: 1573-6857
    Keywords: Anopheles albimanus ; malaria vector ; transformation ; white gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have cloned and characterized the white gene of Anopheles albimanus. Comparison of the deduced amino acid sequence of this white gene with its homologs from six species of Diptera show that the An. albimanus gene is most similar to the white gene of An. gambiae (92% identity). A spontaneous white-eyed mutant An. albimanus was caused by an approximately 10 kb insertion into a CT dinucleotide repeat region of intron 2 of the white locus. The flanks of this insertion are long (at least 400 bp), nearly perfect inverted terminal repeat sequences. This cloned white gene should be useful as a marker for germ line transformation of An. albimanus.
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  • 69
    ISSN: 1572-9818
    Keywords: Agrobacterium ; Populus ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In recent years, Populus species have acquired an important place in basic and applied research of woody plants. The practical role of Populus species in world forestry and their importance to research as a woody-plant model have led to increasing interest in tissue-culture and molecular techniques, as well as the development of transformation procedures for this genus. A simple technical procedure is described here step-by-step, for the first time, as a routine method for transforming Populus tremula using a disarmed Agrobacterium tumefaciens hypervirulent strain. The procedure begins with the inoculation of stem explants with bacterial suspension, followed by a short period of co-cultivation on a highly regenerative medium. Transformed shoots are selected on regeneration medium containing antibiotics and the presence of the inserted target genes is checked using a rapid and efficient PCR test. Selected shoots are transferred to a rooting medium, under the same selection pressure, and propagated via stem cuttings. Selected plants can be hardened and transferred to the green-house within 4 months of inoculation. The method has proven efficient for several gene constructs, selection on Kan or Hyg, and three different Agrobacterium strains.
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  • 70
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    Plant molecular biology reporter 15 (1997), S. 209-215 
    ISSN: 1572-9818
    Keywords: selectable marker ; cereals ; leaf assay ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A very simple leaf assay is described that rapidly and reliably identifies transgenic plants expressing the hygromycin resistance gene, hph or the phosphinothricin resistance gene, bar. Leaf tips were cut from plants propagated either in the glasshouse or in tissue culture and the cut surface embedded in solid medium containing the appropriate selective agent. Non-transgenic barley or rice leaf tips had noticeable symptoms of either bleaching or necrosis after three days on the medium and were completely bleached or necrotic after one week. Transgenic leaf tips remained green and healthy over this period. This gave unambiguous discrimination between transgenic and non-transgenic plants. The leaf assay was also effective for dicot plants tested (tobacco and peas).
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  • 71
    ISSN: 1572-9818
    Keywords: Junction sequence ; transformation ; Agrobacterium tumefaciens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genomic integration of transferred T-DNA is traditionally analyzed by Southern hybridization; however, these analyses often do not provide sufficient information pertaining to the transformation event. Analysis of the junction sequences spanning the region between the T-DNA borders and plant genomic DNA, give a clear demonstration of genomic integration. The procedures available for border junction analysis can be problematic, therefore a simplified method was developed for plants transformed by Agrobacterium tumefaciens harboring the binary vector with pBI121 backbone.
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  • 72
    ISSN: 1573-5044
    Keywords: Agrobacterium ; coat protein ; grapevine ; hairy root ; nepovirus ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hairy root cultures of grapevine were obtained from plantlets co-inoculated by virulent Agrobacterium rhizogenes strains and disarmed A. tumefaciens strains harbouring the binary vectors pKHG4 and pKVHG 2+. These plasmids contain the nptII, hpt and gus genes and differ for the presence of the gene encoding for the grapevine chrome mosaic virus coat protein. For the cultivar ‘Gravesac’, 72% of the excised root tips initiated hairy root cultures on growth regulator-free media. According to the nature of the strains used in co-inoculation, co-transformation frequencies of the hairy root clones ranged from 4 to 16%. Co-transformed roots showed resistance to kanamycin and hygromycin but responses varied from clone to clone. Fluorometric GUS expression and GCMV coat protein production showed a large variability among hairy root clones co-transformed by pKHVG2+. Though the presence of gus, nptII and GCMV coat protein genes was checked by polymerase chain reaction and Southern blotting, it was difficult to establish a clear relationship between expression of the different transgenes. The regeneration of plants was not achieved, but the possibility to graft in vitro transgenic roots to non transformed shoot systems could permit rapid testing of the resistance induced by nepovirus coat protein in roots of cultivars that are recalcitrant to A. tumefaciens-mediated transformation.
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  • 73
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    Euphytica 96 (1997), S. 153-161 
    ISSN: 1573-5060
    Keywords: cassava ; transformation ; review ; somatic embryogenesis ; adventitious shoot formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A prerequisite for the development of a successful transformation system is the availability of efficient regeneration systems. Up to 1995 the only available regeneration system in cassava was an organized type of somatic embryogenesis. Transformation of these organized somatic embryogenic cultures with particle bombardment or Agrobacterium tumefaciens resulted in chimeric transformed embryos. However, the transformed sector was lost after repeated cycles of secondary somatic embryogenesis. After 1995 a less organized system of somatic embryogenesis was developed, so called friable embryogenic callus (FEC) and a system of adventitious shoot regeneration. The FEC regeneration system was combined successfully with particle bombardment. Selection of transgenic plants was based on either luciferase activity, or resistance to the aminoglycoside paromomycin or the herbicide phosphinothricin. Furthermore, protoplasts of FEC are able to regenerate into plants and can be transformed by electroporation. The adventitious shoot regeneration system was combined successfully with Agrobacterium tumefaciens. For this mature somatic embryos were cocultivated with Agrobacterium and cultured for adventitious shoot development. After selection based on the aminoglycoside geneticin or on hygromycin transgenic plants were formed.
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  • 74
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    Euphytica 96 (1997), S. 167-172 
    ISSN: 1573-5060
    Keywords: Coffea spp. ; coffee ; genetic ; in vitro ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The most important advances obtained on in vitro coffee regeneration systems and in coffee genetic transformation, drawing perspectives and scopes to further studies in these fields are presented and discussed.
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  • 75
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    Plant molecular biology 35 (1997), S. 205-218 
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; rice ; transformation ; monocotyledons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Agrobacterium tumefaciens has been routinely utilized in gene transfer to dicotyledonous plants, but monocotyledonous plants including important cereals were thought to be recalcitrant to this technology as they were outside the host range of crown gall. Various challenges to infect monocotyledons including rice with Agrobacterium had been made in many laboratories, but the results were not conclusive until recently. Efficient transformation protocols mediated by Agrobacterium were reported for rice in 1994 and 1996. A key point in the protocols was the fact that tissues consisting of actively dividing, embryonic cells, such as immature embryos and calli induced from scutella, were co-cultivated with Agrobacterium in the presence of acetosyringonc, which is a potent inducer of the virulence genes. It is now clear that Agrobacterium is capable of transferring DNA to monocotyledons if tissues containing ‘competent’ cells are infected. The studies of transformation of rice suggested that numerous factors including genotype of plants, types and ages of tissues inoculated, kind of vectors, strains of Agrobacterium, selection marker genes and selective agents, and various conditions of tissue culture, are of critical importance. Advantages of the Agrobacterium-mediated transformation in rice, like on dicotyledons, include the transfer of pieces of DNA with defined ends with minimal rearrangements, the transfer of relatively large segments of DNA, the integration of small numbers of copies of genes into plant chromosomes, and high quality and fertility of transgenic plants. Delivery of foreign DNA to rice plants via A. tumefaciens is a routine technique in a growing number of laboratories. This technique will allow the genetic improvement of diverse varieties of rice, as well as studies of many aspects of the molecular biology of rice.
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  • 76
    ISSN: 1573-5028
    Keywords: CaMV 35S promoter ; Catharanthus roseus ; FMV 34S promoter ; particle bombardment ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Activities of several CaMV 35S and FMV 34S promoter derivatives fused to the gusA reporter gene were compared in suspension-cultured Catharanthus roseus cells that were transiently and stably transformed using particle bombardment. Our data demonstrate that the 35S and a deletion derivative of the 34S promoter combined with particle bombardment form useful tools for genetic engineering of C. roseus cells. Our results disagree on several points with activities of 35S and 34S promoter derivatives reported for tobacco, indicating that absolute and relative promoter activities can differ between plant species.
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  • 77
    ISSN: 1573-5028
    Keywords: competitive PCR ; flavonoid pathway ; Forsythia ; gene expression ; transformation ; woody ornamentals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression, during flower development, of the gene encoding the anthocyanin pathway key enzyme dihydroflavonol 4-reductase (DFR) was investigated in floral organs of Forsythia × intermedia cv. ‘Spring Glory’. Full-length DFR and partial chalcone synthase (CHS) cDNAs, the gene of interest and a flavonoid pathway control gene respectively, were obtained from petal RNA by reverse transcription PCR. Whereas for CHS northern blot analysis enabled the study of its expression pattern, competitive PCR assays were necessary to quantify DFR mRNA levels in wild-type plants and in petals of 2 transgenic clones containing a CaMV 35S promoter-driven DFR gene of Antirrhinum majus. Results indicated a peak of CHS and DFR transcript levels in petals at the very early stages of anthesis, and different expression patterns in anthers and sepals. In comparison to wild-type plants, transformants showed a more intense anthocyanin pigmentation of some vegetative organs, and a dramatic increase in DFR transcript concentration and enzymatic activity in petals. However, petals of transformed plants did not accumulate any anthocyanins. These results indicate that other genes and/or regulatory factors should be considered responsible for the lack of anthocyanin production in Forsythia petals.
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  • 78
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    Plant cell, tissue and organ culture 49 (1997), S. 71-74 
    ISSN: 1573-5044
    Keywords: cocultivation ; monocot ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Root explants from in vitro-grown plants of Anthurium andraeanum ‘Alii’ and Anthurium interspecific hybrid ‘UH1060’ readily produced multiple shoots under weak light on a modified Murashige & Skoog medium containing 2.2 μM BA. Regenerated ‘UH1060’ plants grew normally and flowered within 16 months after transfer to the greenhouse. Cocultivation of root cuttings with Agrobacterium tumefaciens LBA4404 carrying the binary vector pCa2Att resulted in kanamycin-resistant ‘Anuenue’ shoots transgenic for neo and att and recovered more than one year after culture on selection media. Transformation efficiency (number of explants with transgenic shoots per total explants) was 1.3%. Other cultivars (‘UH1003’, ‘UH1060’, ‘Rudolph’, and ‘Mauna Kea’) failed to produce shoots under the transformation condition employed.
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  • 79
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    Water, air & soil pollution 95 (1997), S. 35-43 
    ISSN: 1573-2932
    Keywords: humic acid ; mercury ; transport ; transformation ; soil-plant system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The influence of humic acid (HA) on the transport and transformation of mercury (Hg) in soil was studied. No available Hg could be detected (〈2.5 μg kg-1) in alluvial soil when the content of HA-carbon (HA-C) was higher than 0.2 g kg-1 although a large amount of Hg (8 μg kg-1) was applied to the soil. The available Hg decreased with the increase of HA in purple soil (r=0.735). There are significant correlations between HA concentration and organic Hg in the tested soils (r=0.974 for the purple soil and r=0.979 for the alluvial soil). The increase of HA results in decrease of Hg absorbed by plant from the soil. A loss of Hg from soil caused by microbes was observed.
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  • 80
    ISSN: 1573-2932
    Keywords: desorption ; ferrihydrite ; modelling ; phosphate ; TEM ; transformation ; wastewater
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The presence of phosphate retards the transformation of ferrihydrite into crystalline products. Increasing phosphate from 0 to 1 mole % results in an order of magnitude decrease in the rate of transformation of ferrihydrite at pH 12. Levels of phosphate of ∼1 mol % suppress the formation of goethite (α-FeO(OH)) and result in the formation of a product consisting of η-Fe2O3. Higher levels of phosphate result in the ferrihydrite remaining amorphous, even after several hundred hours. Phosphate prevents formation of goethite by hindering the dissolution of ferrihydrite rather than by interfering with nucleation and growth of goethite in solution. The transformation rate of pure ferrihydrite is also strongly inhibited in the presence of dissolved phosphate. This is due to surface complexation. The transformation rate was measured at temperatures of 60 °C and 70 °C. The rate of transformation was found to be described by either (i) a solid-state reaction equation for powdered compacts or (ii) a zero-order reaction controlled by desorption. The transformation of the ferrihydrite matrix was accompanied by the loss of the phosphate trace component. X-ray diffraction indicates that no solid solution involving phosphate substitution into η-Fe2O3 is formed. Transmission electron microphotographs of the original precipitates containing phosphate confirm the presence of the phosphate and demonstrate its involvement in linking together extremely small particles of ferrihydrite.
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  • 81
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    Water, air & soil pollution 95 (1997), S. 35-43 
    ISSN: 1573-2932
    Keywords: humic acid ; mercury ; transport ; transformation ; soil-plant system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The influence of humic acid (HA) on the transport and transformation of mercury (Hg) in soil was studied. No available Hg could be detected (〈2.5 μg kg−1) in alluvial soil when the content of HA-carbon (HA-C) was higher than 0.2 g kg−1 although a large amount of Hg (8 μg kg−1) was applied to the soil. The available Hg decreased with the increase of HA in purple soil (r=0.735). There are significant correlations between HA concentration and organic Hg in the tested soils (r=0.974 for the purple soil and r=0.979 for the alluvial soil). The increase of HA results in decrease of Hg absorbed by plant from the soil. A loss of Hg from soil caused by microbes was observed.
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  • 82
    ISSN: 1572-8773
    Keywords: NMR and EPR spectroscopy ; speciation ; transformation ; vanadate ; vanadyl ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Three strains of Saccharomyces cerevisiae, SC-1, DBVPG 6173 and DBVPG 6037, were studied for vanadate resistance in complex Sabouraud medium since they did not thrive in different minimal media (yeast nitrogen base with and without amino acids). The strain SC-1 was resistant up to 16 mm of vanadate, whereas the strains DBVPG 6173 and DBVPG 6037 were inhibited by 8 mm and 4 mm vanadate, respectively. The vanadate resistance in strain SC-1 was constitutive and due to the reduction of this oxyanion to vanadyl, which was detected by EPR spectroscopy and visible spectroscopy. The transformation of vanadate to vanadyl took place during the exponential growth phase; 10 mm of vanadate was reduced to vanadyl outside the cells since the oxyanion was not detected in the cell biomass and only a negligible concentration of vanadyl (25 nmoles mg cells dry weight) was found in the biomass. The other two vanadate-sensitive yeast strains only accumulated vanadate and did not reduce the oxyanion to vanadyl.
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  • 83
    ISSN: 1573-2932
    Keywords: desorption ; ferrihydrite ; modelling ; phosphate ; TEM ; transformation ; wastewater
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The presence of phosphate retards the transformation of ferrihydrite into crystalline products. Increasing phosphate from 0 to 1 mole % results in an order of magnitude decrease in the rate of transformation of ferrihydrite at pH 12. Levels of phosphate of ∼1 mol % suppress the formation of goethite (α-FeO(OH)) and result in the formation of a product consisting ofη-Fe2O3. Higher levels of phosphate result in the ferrihydrite remaining amorphous, even after several hundred hours. Phosphate prevents formation of goethite by hindering the dissolution of ferrihydrite rather than by interfering with nucleation and growth of goethite in solution. The transformation rate of pure ferrihydrite is also strongly inhibited in the presence of dissolved phosphate. This is due to surface complexation. The transformation rate was measured at temperatures of 60 °C and 70 °C. The rate of transformation was found to be described by either (i) a solid-state reaction equation for powdered compacts or (ii) a zero-order reaction controlled by desorption. The transformation of the ferrihydrite matrix was accompanied by the loss of the phosphate trace component. X-ray diffraction indicates that no solid solution involving phosphate substitution intoη-Fe2O3 is formed. Transmission electron microphotographs of the original precipitates containing phosphate confirm the presence of the phosphate and demonstrate its involvement in linking together extremely small particles of ferrihydrite.
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  • 84
    ISSN: 1573-4919
    Keywords: endothelin ; fibroblasts ; receptors ; transformation ; oncogene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Endothelin-1 (ET-1), a peptide isolated from the culture medium of endothelial cells, mediates a variety of physiological and pathological responses including mitogenesis. We have compared the expression of ET receptors in untransformed versus ras-transformed NIH-3T3 murine fibroblasts and in untransformed versus SV40-transformed Wl38 (VA13) human fibroblasts by ligand binding and Northern analysis. NIH-3T3 and Wl38 cells displayed high affinity (200 and 220 pM) and high density (23,000 sites/cell and 14,000 sites/cell for NIH-3T3 and Wl38 cells, respectively) ET receptors. Competition binding experiments using subtype-selective ligands identified these receptors as the ETA subtype. Addition of ET-1 to the cells produced a concentration-dependent increase in intracellular calcium release. Both ras-transformed NIH-3T3 cells and SV40-transformed Wl38 cells (VA13) completely lacked [125I]ET-1 binding and failed to release calcium when exposed to ET-1. Northern analysis of the polyadenylat ed RNA (polyA RNA) isolated from untransformed and transformed cells revealed that the steady-state level of ETA receptor RNA was 90-95% less in transformed cells compared to untransformed cells. Thus, the loss of ET receptors as well as the receptor-mediated responses in transformed cells can be explained by down-regulation of ET receptor mRNA.
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  • 85
    ISSN: 1572-9788
    Keywords: adventitious root formation ; Agrobacterium tumefaciens ; A. rhizogenes ; rootstock ; rose ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Transgenic plants of the rootstock Rosa hybrida L. cv. Moneyway were produced via a two-step procedure. First, kanamycin-resistant roots were generated on stem slices from micropropagated shoots, which were cocultivated with Agrobacterium tumefaciens containing the neomycin phosphotransferase II (NPTII) gene for conferring kanamycin resistance, together with individual ROL genes from A. rhizogenes. Root formation was quite efficient and up to two kanamycin-resistant roots per stem slice were produced. In the second step, these roots were used to regenerate transgenic plants via somatic embryogenesis. Although regeneration lasted up to 12 months, production of several transformants was successfully accomplished. Untransformed escapes were not found, indicating that the initial selection on kanamycin resistance was reliable. The presence of a combination of ROLA, B and C genes enhanced adventitious root formation on micropropagated shoots and explants of stems and leaves. It appears that the auxin sensitivity was increased to such a degree that cells were able to respond even to endogenous auxins present in shoots and leaves. Rooting experiments in greenhouse demonstrated that adventitious root formation on cuttings was improved threefold upon introduction of these ROL genes. It is concluded that a method was developed for the production of ROL gene transformed roses with improved rooting characteristics.
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  • 86
    ISSN: 1572-9788
    Keywords: Agrobacterium tumefaciens ; bar gene ; grain legume ; Lupinus angustifolius ; phosphinothricin ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Transgenic plants of Lupinus angustifolius L. (cvs. Unicrop and Merrit) were routinely generated using Agrobacterium-mediated gene transfer to shoot apices. The bar gene for resistance to phosphinothricin (PPT, the active ingredient of the herbicide Basta) was used as the selectable marker. After co-cultivation, the shoot apex explants were transferred onto a PPT-free regeneration medium and their tops were thoroughly wetted with PPT solution (2 mg/ml). The multiple axillary shoots developing from the shoot apices were excised onto a medium containing 20 mg/l PPT. The surviving shoots were transferred every second week onto fresh medium containing 20 mg/l PPT. At each transfer, the number of surviving shoots decreased, until it stabilized. Indeed, some of these chimeric shoots surviving the PPT selection, eventually produced new green healthier axillary shoots which could be transferred to soil. This whole process took from 5 to 9 months after co-cultivation. Average transformation frequencies of 2.8% for cv. Unicrop and of 0.4% for the commercial cultivar Merrit were achieved. Molecular analysis of T0, T1, and T2 generations demonstrated stable integration of the foreign gene into the plant genome and expression of the integrated gene. Transformed plants of the T1 and T2 generations were resistant in glasshouse trials where the herbicide Basta (0.1 mg/ml) was sprayed onto whole plants. These results demonstrate that Agrobacterium-mediated gene transfer to preorganised meristematic tissue combined with axillary regeneration can form the basis of a routine transformation system for legume crop species which are difficult to regenerate from other explants.
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  • 87
    ISSN: 1572-9788
    Keywords: Bacillus thuringiensis ; Chilo suppressalis ; biolistic ; rice ; Scirpophaga incertulas ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Rice cultivars of isozyme group V include high-quality, aromatic rices that are difficult to improve by traditional methods because of the loss of quality characters upon sexual hybridization. Their low-tillering plant type predisposes them to economic loss from attack by stem borers, a group of insects to which they are susceptible. We report here the enhancement of stem borer resistance in cv. Tarom Molaii through transformation by microprojectile bombardment. Embryogenic calli derived from mature seeds were bombarded with gold particles coated with plasmid pCIB4421, carrying a synthetic truncated toxin gene based on the cryIA(b) gene from Bacillus thuringiensis, and plasmid pHygII, carrying the hygromycin phosphotransferase (hpt) selectable marker gene. Inclusion of 50 mg/l hygromycin B in culture media from bombardment through to rooting of plantlets eliminated escapes. The procedure generated three independent hpt transformants of which two also contained the cryIA(b) gene. One such line (No. 827) produced truncated (67 kDa) CryIA(b) protein equivalent to about 0.1% of total soluble protein. The cryIA(b) gene was controlled by the promoter of the maize C4 PEP carboxylase gene and was expressed in leaf blades but was not expressed to a detectable level in dehulled mature grain. Line 827 contained about 3 copies of the cryIA(b) gene which segregated as a single dominant Mendelian locus in the second (T1) and third (T2) generations and co-segregated with enhanced resistance to first-instar larvae of striped stem borer (Chilo suppressalis) and yellow stem borer (Scirpophaga incertulas). T2 line 827-6 homozygous for the cryIA(b) gene showed no dead hearts or whiteheads after infestation with stem borers, whereas T2 line 827-25 lacking the gene averaged 7 dead hearts per plant and 2.25 whiteheads per plant. These results establish that transformation of high-quality rices of group V is a feasible alternative to sexual hybridization.
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  • 88
    ISSN: 1573-9368
    Keywords: β-GUS ; embryogenesis reinduction ; kanamycin ; NPTII ; Piceaglauca ; selection ; somatic embryos ; somaticseedlings ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transformed white spruce cultures containing immature Stage I somatic embryos, were developed after particle bombardment of somatic embryos with pBI 426, carrying an expression cassette with a gus::nptII fusion gene. These Stage I cultures did not show tolerance to kanamycin concentrations above 3 to 5 mg l−1, although assays for GUS and NPTII showed that functional enzymes were present in the transgenic tissue. Embryonic liquid suspension cultures were initiated from this transformed tissue. After treatment on agar-solidified maturation medium with 48 μm (±)-ABA high numbers of Stage III (cotyledonary) somatic embryos were produced. When subjected to an embryogenesis re-induction process with 2,4-D and BA, these Stage III embryos produced a new generation of Stage I embryogenic tissues which could tolerate 5--10 mg l−1 kanamycin. Stage III somatic embryos could alternatively be placed onto germination medium for the development of somatic seedlings. When germinated in the presence of 20 mg l−1 kanamycin, 77% of inoculants were resistant. The stability of integration of the gus::nptII fusion gene in the genome of white spruce Stage III somatic embryos and somatic seedlings was confirmed through Southern blot analysis
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  • 89
    ISSN: 1573-9368
    Keywords: Ac ; Agrobacterium ; aspen ; Populus ; transformation ; transposition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Aspen (Populus tremula) and hybrid aspen (P. tremula × P. tremuloides) were transformed with different gene constructs using two types of promoter. The aim was to determine the influence of the reporter gene rolC, controlled by promoters of viral or plant origin, on genetic and morphologic expression of different transgenic aspen clones. An improved transformation method using leaf discs was developed, by which putative transgenic plantlets were regenerated at high efficiencies (up to 34%) on kanamycin-containing medium. Transgenic aspen carrying the rolC gene from Agrobacterium rhizogenes under control of the cauliflower-35S-promoter are reduced in size with smaller leaves, whereas aspen transgenic for the same rolC gene, but under control of the light inducible rbcS promoter from potato, are only slightly reduced in size compared to untransformed controls. However, all clones carrying 35S-rolC and rbcS-rolC genes revealed light-green colouration of leaves when compared to untransformed aspen. Owing to this special feature, constructs were used in which expression of the rolC gene was inhibited by insertion of a transposable element, Ac, from maize. Transgenic aspen transformed with the 35S-Ac-rolC and rbcS-Ac-rolC genes were morphologically similar to untransformed aspen, but out of 54 independently regenerated 35S-Ac-rolC transgenic aspen clones, 30 clones showed light-green/dark green variegated leaves. In contrast, out of 19 independently transformed rbcS-Ac-rolC aspen clones, only two clones revealed light-green/dark green variegated leaves. The role of bacterial strains in transformation, and molecular genetics of transgenic aspen plants (including the function of the transposable element, Ac, in the aspen genome) are discussed
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  • 90
    ISSN: 1573-9368
    Keywords: transformation ; Erysiphe graminis ; powderymildew ; coleoptile
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A transient assay is described that should allow evaluation of the role of host genes in disease response by enhancing or disrupting expression of those genes in specific cells and looking for effects on disease development. The assay also has the potential for assessing utility of host and non-host genes in enhancing resistance to disease in transgenic plants. Particle bombardment with a helium discharge particle gun was utilized to transiently express genes in epidermal cells of coleoptiles of barley (Hordeum vulgare). An anthocyanin reporter gene construct provided a means of identifying those cells that were transiently expressing introduced DNA. Optimal transient expression rates were achieved two days following bombardment with 1800 psi helium pressure, 1.0 μm diameter gold particles, and coleoptile pre- and post-treatment in 0.30--0.35 m mannitol/sorbitol. Under optimal conditions, at least 35 cells expressed anthocyanin per bombardment. Transiently expressing cells were inoculated with the fungal pathogen, Erysiphe graminis f. sp. hordei, and fungal development observed. Neither the bombardment procedures, the presence of nearby dead cells, nor accumulation of anthocyanin within living cells affected fungal development in living cells. Therefore, incorporation of disease-related genes onto the same plasmid as the reporter genes will allow evaluation of the role of those genes in disease development or suppression. Since particle bombardment is possible with a great range of different plant tissues, the described methodology should exhibit wide applicability for evaluating genes in diverse plant-pathogen interactions, as well as genes involved in many other biological processes
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  • 91
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    Transgenic research 6 (1997), S. 279-288 
    ISSN: 1573-9368
    Keywords: Agrobacterium tumefaciens ; Brassica napus ; wintercultivar ; transformation ; GUS ; hygromycin ; kanamycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An efficient protocol for Agrobacterium tumefaciens-mediated transformation of six commercial Brassica napus winter cultivars is described. Two B. napus spring cultivars were analysed for comparison. Five strains of A. tumefaciens with different combinations of nopaline and octopine chromosomal backgrounds and virulence plasmids were used for cocultivation. Selection of putative regenerated transgenic plants was performed on kanamycin- or hygromycin-containing media. The scores of transgenic plants were calculated on the basis of GUS (β-glucuronidase) activity, detected by the histochemical X-Gluc test. Target tissue derived from the cut surface of cotyledon petioles resulted in successful transformation with all the winter cultivars tested. Target tissue from hypocotyl segments resulted in a successful transformation with only one winter cultivar. The transformation rates for B. napus winter cultivars in this study were higher than in previous reports. Southern blot analysis revealed that integration of marker genes occurred in single and in multiple copies and at multiple loci in the genome. The transgenic plants all grew normally and developed fertile flowers after a vernalization period. After self-pollination, Southern blot analysis of selected GUS active F1 plants revealed that introduced marker genes were stably inherited to the next generation. These data demonstrate that morphologically normal, fertile transgenic plants of B. napus winter cultivars can be achieved with both nopaline- and octopine-derived A. tumefaciens strains. This protocol should have a broad application in improvement of Brassica napus winter cultivars by introduction of foreign genes
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  • 92
    ISSN: 1573-9368
    Keywords: peanut ; engineered virusresistance ; tomato spotted wiltvirus ; transformation ; nucleocapsid ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucleocapsid gene of tomato spotted wilt virus Hawaiian L isolate in a sense orientation, and the GUS and NPTII marker genes, were introduced into peanut (Arachis hypogaea cv. New Mexico Valencia A) using Agrobacterium-mediated transformation. Modifications to a previously defined transformation protocol reduced the time required for production of transformed peanut plants. Transgenes were stably integrated into the peanut genome and transmitted to progeny. RNA expression and production of nucleocapsid protein in transgenic peanut were observed. Progeny of transgenic peanut plants expressing the nucleocapsid gene showed a 10- to 15-day delay in symptom development after mechanical inoculations with the donor isolate of tomato spotted wilt virus. All transgenic plants were protected from systemic tomato spotted wilt virus infection. Inoculated non-transformed control plants and plants transformed with a gene cassette not containing the nucleocapsid gene became systemically infected and displayed typical tomato spotted wilt virus symptoms. These results demonstrate that protection against tomato spotted wilt virus can be achieved in transgenic peanut plants by expression of the sense RNA of the tomato spotted wilt virus nucleocapsid gene
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  • 93
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    Transgenic research 6 (1997), S. 329-336 
    ISSN: 1573-9368
    Keywords: Agrobacterium ; SAAT ; sonication ; transformation ; wounding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plant transformation via Agrobacterium can be limited by both host specificity and the inability of Agrobacterium to reach the proper cells in the target tissue. Described here is a new and efficient Agrobacterium-based transformation technology that overcomes these barriers and enhances DNA transfer in such diverse plant groups as dicots, monocots, and gymnosperms. This new technology, called sonication-assisted Agrobacterium-mediated transformation (SAAT), involves subjecting the plant tissue to brief periods of ultrasound in the presence of Agrobacterium. Scanning electron and light microscopy reveal that SAAT treatment produces small and uniform fissures and channels throughout the tissue allowing the Agrobacterium easy access to internal plant tissues. Unlike other transformation methods, this system has the potential to transform meristematic tissue buried under several cell layers. SAAT increases transient transformation efficiency in several different plant tissues including leaf tissue, immature cotyledons, somatic and zygotic embryos, roots, stems, shoot apices, embryogenic suspension cells and whole seedlings. A 100- to 1400-fold increase in transient β- glucuronid ase expression has been demonstrated in various tissues of soybean, Ohio buckeye, cowpea, white spruce, wheat and maize. Stable transformation of both soybean and Ohio buckeye has been obtained using SAAT of embryogenic suspension culture tissues. For soybean, SAAT treatment was necessary to obtain stable transformation with this tissue
    Type of Medium: Electronic Resource
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  • 94
    ISSN: 1573-9368
    Keywords: transgenic cotton ; bialaphos ; particle bombardment ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Resistance to bialaphos, a non-selective herbicide, was intro duced into cotton through genetic engineering. A gene encoding phosphinothric in acetyltransferase (bar) from Streptomyces hygroscopicus was inserted into elite varieties of cotton through particle bombardment. Based on the marker gene, β-glucuronidase (gus) expression, a total of 18 Pima (Gossypium barbadense), 45 DP50 (G. hirsutum L.), 20 Coker 312 (G. hirsutum) and 2 El Dorado (G. hirsutum) transgenic plants were recovered. Integration of the bar gene into cotton genomic DNA was confirmed by Southern blot analysis and gene expression was confirmed by northern blot and enzyme assays. Herbicide (Basta®) tolerance up to 15 000 ppm was demonstrated in greenhouse trials. The newly introduced herbicide tolerance trait is inherited in a Mendelian fashion in the progenies of germline transformants. This study demonstrates the potential for particle bombardment to introduce commerically important genes directly into elite varieties of cotton. This mode of gene transfer can expedite the introduction of transgenic cotton products into world markets
    Type of Medium: Electronic Resource
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  • 95
    ISSN: 1573-9368
    Keywords: GUS ; matrix attachment regions ; Populus ; transformation ; transgene expression ; Agrobacterium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We tested the value of a matrix attachment region (MAR) fragment derived from a tobacco gene for increasing the frequency of Agrobacterium-mediated transformation. A binary vector that carried a GUS reporter gene containing an intron and an nptII gene was modified to contain flanking MAR elements within the T-DNA borders. Vectors containing or lacking MARs were then used to transform tobacco, a readily transformabl e poplar clone (Populus tremula × P. alba), and a recalcitrant poplar clone (Populus trichocarpa × P. deltoides). MARs increased GUS gene expression approximately 10-fold in the two hybrid poplar clones and twofold in tobacco one month after cocultivation with Agrobacterium; MARs also increased the frequency of kanamycin-resistant poplar shoots recovered
    Type of Medium: Electronic Resource
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  • 96
    Electronic Resource
    Electronic Resource
    Springer
    Journal of porous materials 4 (1997), S. 165-169 
    ISSN: 1573-4854
    Keywords: ferrihydrite ; hematite ; BET ; porosity ; catalyst ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Abstract Ferrihydrite is a poorly-ordered iron oxyhydroxide which possesses a high specific surface area (〉200 m2/g). This makes it an important constituent for surface related processes in nature and catalysis. This study focuses on the precautions that have to be taken in the sample preparation prior to BET measurements in order to avoid essential structural transformations. The experiments showed that temperatures between 90 and 120°C are necessary to remove surface contaminations and simultaneously prevent structural transformation of the ferrihydrite during the outgassing procedure. Furthermore, care has to be taken to keep the humidity low in the immediate proximity of the sample, due to the strong impact of this parameter on the structural stability of ferrihydrite.
    Type of Medium: Electronic Resource
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  • 97
    Electronic Resource
    Electronic Resource
    Springer
    Acta mechanica solida Sinica 9 (1996), S. 357-363 
    ISSN: 0894-9166
    Keywords: smart composites ; shape memory effect ; transformation ; constitutive relations ; toughening ; thermomechanics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics , Physics
    Notes: Abstract The smart composite materials reinforced by SMA show a high performance and special deformation behavior. The thermomechanical constitutive formulas of the composites are derived by means of Eshelby's equivalent inclusion method and Mori-Tanaka's mean field concept. The interaction between the inclusion and crack and toughening mechanism are considered and the energy release rate of a crack in the smart composite is calculated. This work shows that there are the multiple mechanisms contributing to the toughening of the smart composite materials reinforced by SMA.
    Type of Medium: Electronic Resource
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  • 98
    ISSN: 1573-6857
    Keywords: alcohol dehydrogenase ; Drosophila ; enhancer ; regulation ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A region (NS1) that acts like an enhancer is located approximately 300 bp upstream of the larval cap site in theAdh gene ofD. melanogaster. When this sequence is deleted (ΔNS1), the gene fails to express ADH protein. Gene expression can be restored by placing a secondAdh gene with an intact enhancer elsewhere on the same plasmid. In these circumstances, both genes are expressed equally regardless of their orientation on the plasmid. In this report we further characterize the interactions that occur when a single enhancer activates expression from a proximal and distant promoter. We have made the following observations: (1) While the two genes are expressed equivalently, their expression relative to a plasmid carrying two intact genes is reduced by a factor of 2 to 6 depending on the orientation of the two genes. (2) The single enhancer drives expression of both genes on any given plasmid molecule. (3) The enhancer does not interact with theAdh gene from which the NS7 region (which spans the larval TATA box) is removed. (4) Expression of the ΔNS1 gene can be restored by an intact gene when both are inserted together into theDrosophila genome via P element-mediated transformation. (5) Increasing the separation between the two genes on a plasmid by up to 15 kbp does not prevent the restoration of expression of the ΔNS1 gene. We propose a model that explains how a single enhancer can stimulate equal expression from two genes.
    Type of Medium: Electronic Resource
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  • 99
    Electronic Resource
    Electronic Resource
    Springer
    Apoptosis 1 (1996), S. 126-130 
    ISSN: 1573-675X
    Keywords: SV40 ; tissue transglutaminase ; transformation ; tumour marker
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract During our search for novel transformation-sensitive proteins whose synthesis is abolished in tumour cells we found a cDNA clone coding for tissue transglutaminase. This enzyme was identified, at the protein as well as the mRNA level, in normal human fibroblasts, but was completely missing in their matched SV40 transformed counterparts. Since tissue transglutaminase has been implicated in cell cycle regulation and apoptosis, we investigated the possibility of whether this enzyme might represent a negative marker for tumour cells. We found that its synthesis varied largely among 10 cell lines derived from spontaneous mesenchymal tumours. While cells from a rhabdomyosarcoma and a chondrosarcoma did not produce it at all, an extremely high expression was observed in cells from an osteosarcoma and a liposarcoma. Thus, tissue transglutaminase is not a tumour-related marker.
    Type of Medium: Electronic Resource
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  • 100
    Electronic Resource
    Electronic Resource
    Springer
    Zeitschrift für angewandte Mathematik und Physik 47 (1996), S. 821-836 
    ISSN: 1420-9039
    Keywords: Stream functions ; vector potential ; transformation ; vortex flows
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mathematics , Physics
    Notes: Abstract Introducing a vector potential, that is based on a pair of stream functions, and a velocity potential, antisymmetric equations for the stream functions are derived with the help of a variational principle. It is found that the equations are in a suitable form to investigate flows with helical symmetry, and, for example, to connect upstream axisymmetric flows with downstream helical flows. The special case of a transition from an upstream solid-body vortex to a downstream helical flow is investigated in detail. Furthermore, the stream-function equations are particularly useful to investigate general small-amplitude inertia waves on vortex flows. Time-dependent helical flows that are time-independent in a suitably rotating frame of reference can also be discussed with the proposed method.
    Type of Medium: Electronic Resource
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