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  • 1990-1994  (39)
  • 2,4-dichlorophenoxyacetic acid (2,4-D)
  • DGGE
  • Information technology, Management
  • Recombinant DNA
  • bioremediation
  • 1
    ISSN: 1432-0827
    Keywords: Amelogenin ; Expression ; Enamel ; Recombinant DNA ; Tooth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract A mouse cDNA encoding a 180 amino acid amelogenin was subcloned into the pET expression plasmid (Novagen, Madison, WI) for production in Escherichia coli. A simple growth and purification protocol yields 20–50 mg of 95–99% pure recombinant amelogenin from a 4.5-liter culture. This is the first heterologous expression of an enamel protein. The expressed protein was characterized by partial Edman sequencing, amino acid composition analysis, SDS-PAGE, Western blotting, laser desorption mass spectrometry, and hydroxyapatite binding. The recombinant amelogenin is 179 amino acids in length, has a molecular weight of 20,162 daltons, and hydroxyapatite binding properties similar to the porcine 173 residue amelogenin. Solubility analyses showed that the bacterially expressed protein is only sparingly soluble in the pH range of 6.4–8.0 or in solutions 20% saturated with ammonium sulfate. The purified protein was used to generate rabbit polyclonal anti-amelogenin antibodies which show specific reaction to amelogenins in both Western blot analyses of enamel extracts and in immunostaining of developing mouse molars.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of cancer research and clinical oncology 120 (1994), S. 454-464 
    ISSN: 1432-1335
    Keywords: Xeroderma pigmentosum fibroblasts ; DNA repair ; Recombinant DNA ; cDNA libraries ; Phage λgt10 ; pBluescript vector ; In vitro transcripts ; Differential hybridization ; Mitochondrial neuromyopathies ; Genetic defects ; Uracil-DNA glycosylase ; Glyceraldehyde-3-phosphate dehydrogenase ; Lactate dehydrogenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Differential hybridization was used to detect repair defects in xeroderma pigmentosum (XP) that are not amenable to current analyses. cDNA libraries were constructed from cytoplasmic RNA of normal and XP fibroblast strains (complementation groups A and D) and analyzed for differential gene expression. More than 40000 λgt10 cDNA clones were differentially screened with in vitro transcripts made from cDNA in the pBluescript vector. Six differential clones were detected in the libraries of the XP group A and D strains which caused stronger or weaker signals when probed with transcripts from XP strains than with those from the normal strains. Two clones coded for mitochondrial genes: mitochondrial 16 S rRNA and ATPase 6L. Overexpression of mitochondrial genes in XP may indicate that functions of the ATP-generating system are impaired since such functions are intensified whenever they become insufficient, for example as a consequence of DNA damage. It is tempting to assume that abnormal mitochondria are one of the causes for the neurological malfunctions in XP. Furthermore, densitometric analysis of Northern blots revealed that mRNA of lactate dehydrogenase, chain M, was less abundant in four XP group A strains (extent of reduction: 70%) and in two XP group D strains (extent of reduction: 58%). Enzyme activity was also diminished. In addition, mRNA of the gene for glyceraldehyde-3-phosphate dehydrogenase was less expressed in the same XP group A and D fibroblast strains investigated (reduction in both complementation groups: 50%). Both glycolytic enzymes have nuclear functions apart from their role in sugar metabolism. Lactate dehydrogenase, chain M, is identical to a helix-destabilizing protein; it is closely associated with chromatin and unfolded DNA, suggesting a role in DNA synthesis and transcription. The 37-kDa subunit of glyceraldehyde-3-phosphate dehydrogenase is involved in transcription and was shown to be identical to uracil-DNA glycosylase, a base-excision repair enzyme. We presume that the nuclear functions of these glycolytic enzymes may be thwarted in the XP strains investigated and may account for malfunctions in XP, particularly for neurological disturbances.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 243 (1994), S. 253-260 
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Saccharomyces cerevisiae ; Endo-β-glucanase ; Endo-xylanase ; Heterologous expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have developed a method for fast and efficient isolation of enzyme genes from filamentous fungi by combining the ability of Saccharomyces cerevisiae to express heterologous genes with the utilisation of sensitive and reliable enzyme assays. A cDNA library from the fungus Humicola insolens was constructed in a S. cerevisiae/Escherichia coli shuttle vector in E. coli. Sub-pools of the library were subsequently screened for enzyme activity in S. cerevisiae. More than 130 clones were identified as positive in either an endo-β-glucanase or an endo-xylanase assay. Based on a partial characterization of the DNA sequence of the individual clones, they could be grouped into five distinct types of endo-β-glucanases and three types of endo-xylanases. A representative cDNA from each type was sub-cloned in an Aspergillus vector and expressed in A. oryzae. The new cloning method may be an important alternative to traditional cloning methods based on amino acid sequence information.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    World journal of microbiology and biotechnology 10 (1994), S. 472-474 
    ISSN: 1573-0972
    Keywords: Bacillus subtilis ; bioremediation ; copper ; Gram-positive walls
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Purified cell walls from Bacillus subtilis were repeatedly suspended in 5 mm CuCl2 and, after removing unbound Cu, were suspended in 1% (v/v) HNO3 to release bound Cu. The walls were then regenerated by washing in H2O. After five cycles, copper binding actually increased slightly, probably due to enhanced exposure of binding sites in the walls. Thus bacterial walls may be used repeatedly for metal removal during bioremediation of heavy metal pollution.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 43 (1994), S. 521-528 
    ISSN: 0006-3592
    Keywords: carbon tetrachloride ; acetate ; nitrate ; bioremediation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A denitrifying consortium capable of transforming carbon tetrachloride (CCl4) was cultured from aquifer sediment from the U.S. Department of Energy's Hanford Site in southeastern Washington State. To understand the kinetics of the biological destruction of CCl4 by these microbes, a set of experiments, the conditions of which were chosen according to a fractional factorial experimental design, were completed. This article reports on the experimental design along with the results for CCl4, biomass, acetate, nitrate, and nitrite concentrations. These data indicate that growth is inhibited by high nitrite concentrations, whereas CCl4 degradation is slowed by the presence of nitrate and/or nitrite. © 1994 John Wiley & Sons, Inc.
    Additional Material: 5 Ill.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 10 (1994), S. 659-662 
    ISSN: 0749-503X
    Keywords: Recombinant DNA ; purine salvage enzymes ; conserved sequences ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of a Saccharomyces cerevisiae gene encoding a potential adenine phosphoribosyltransferase (APRT) has been determined. The protein encoded by this gene shows a high degree of similarity with APRTs from a variety of other species. The S. cerevisiae gene, named APT2, has been mapped to chromosome IV. The sequence has been deposited in the GenBank data library under Accession Number L14434.
    Additional Material: 2 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Applied Organometallic Chemistry 8 (1994), S. 501-508 
    ISSN: 0268-2605
    Keywords: Amendment ; biomethylation ; bioremediation ; dimethyl selenone ; headspace analysis ; fluorine-induced chemiluminescence ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A possible biological intermediate in the reduction and methylation of selenium oxyanions, dimethyl selenone, was synthesized, and the first experiments involving the amendment of selenium resistant bacterial cultures with this compound are reported. The amount of volatile, reduced selenium-containing species released from these cultures into the headspace is significantly more than that produced in analogous experiments involving sodium selenate amended cultures. Dimethyl selenone is reduced in the presence of dimethyl sulfide and dimethyl disulfide in a complex growth medium, trypticase soy broth with 0.1% nitrate. This reduction occurs whether or not the reduced sulfur compounds are biologically produced.
    Additional Material: 2 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    Journal of Chemical Technology AND Biotechnology 59 (1994), S. 9-23 
    ISSN: 0268-2575
    Keywords: microbial metabolism ; xenobiotics ; biodegradation ; bioremediation ; bioaugmentation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The ability of microorganisms to metabolise xenobiotic compounds has received much attention due to the environmental persistence and toxicity of these chemicals. The microbial degradation of xenobiotics is seen as a cost effective method of removing these pollutants from the environment by a process now known as bioremediation. Microbial treatment of industrial effluents is also possible. Fundamental work has revealed that a wide variety of microorganisms are capable of degrading an equally wide range of organic pollutants. Pure and mixed cultures of microorganisms have been studied and degradation is observed under both aerobic and anaerobic conditions. Breakdown products have been found during work on the degradative pathways involved and toxicological assessments using bacteria and higher organisms (fish, plants) have been used to determine the toxicity of these intermediates. Many of the degradative genes responsible for xenobiotic metabolism are present on plasmids, transposons or are grouped in clusters on chromosomes. This provides clues to the evolution of degradative pathways and makes the task of genetic manipulation easier such that new microbial strains capable of efficiently degrading pollutants can be developed. Several enzymes involved in xenobiotic metabolism have been isolated and factors affecting their activity investigated. Genetically manipulated strains or naturally isolated organisms may be used in the treatment of industrial wastes or as inocula to enhance degradation in the environment. Environmental factors, including pH, temperature, bioavailability, nutrient supply and oxygen availability have been shown to affect xenobiotic biodegradation. These factors must be optimised to obtain a satisfactory microbial treatment process. Using information gained from fundamental research, bioremediation technology has been used to detoxify different contaminated environments and the results of field studies are very encouraging.
    Additional Material: 2 Tab.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Biodegradation 4 (1993), S. 283-301 
    ISSN: 1572-9729
    Keywords: air pollution ; biofiltration ; bioremediation ; bioscrubbing ; off-gas treatment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract This paper gives an overview of present biological techniques for the treatment of off-gases and the techniques that are being developed at the moment. The characteristics, advantages, disadvantages, costs and application area are discussed and compared. Biological off-gas treatment is based on the absorption of volatile contaminants in an aqueous phase or biofilm followed by oxidation by the action of microorganisms. Biofilters, bioscrubbers and biotrickling filters are used for elimination of odour and bioconvertable volatile organic and inorganic compounds and are enjoying increasing popularity. This popularity is a result of the low investment and operational costs involved compared to physico-chemical techniques and the elimination efficiencies that can be obtained. The operational envelop is still extending to higher concentrations and gas flow rates (exceeding 200,000 m3 h−1) and a broader spectrum of degradable compounds. Research and development on the use of membranes and the addition of activated carbon or a second liquid phase to the biological systems may lead to a more efficient elimination of hydrophobic compounds and buffering of fluctuating loads. Shorter adaptation periods can be obtained by inoculation with specialized microorganisms. Improved design and operation are made possible by the growing insights in the kinetics and microbiology and supported by the development of models describing biological off-gas treatment. In conclusion, biotechniques are efficient and cost effective in treating off-gases with concentrations of biodegradable contaminants up to 1–5 g/m3. They could play a justified and important role in air pollution control in the coming years.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 23 (1993), S. 547-548 
    ISSN: 1432-0983
    Keywords: Autonomously replicating sequence ; Recombinant DNA ; Nested deletions ; Mutagenesis ; Schizosaccharomyces pombe
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plasmids pSP1 and pSP2 are two new Schizosaccharomyces pombe ars1 multicopy vectors with the Saccharomyces cerevisiae LEU2 and URA3 genes as selectable markers. They are derivatives of S. cerevisiae integrative plasmids. These plasmids allow classical molecular genetic techniques, such as mutagenesis, nested deletions and sequencing, to be performed directly.
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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 23 (1993), S. 435-442 
    ISSN: 1432-0983
    Keywords: Heat shock ; Recombinant DNA ; Membrane protein ; Nutritional limitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using differential hybridization, a gene preferentially expressed during entry into stationary phase has been isolated. Subsequent analysis indicated that this gene corresponds to a heat-shock gene. The nucleotide sequence has been determined. It revealed a 332 aminoacid protein. No similarities to any previously known protein have been noted. The protein is very hydrophobic and is predicted to have a membraneous localisation. In agreement with this hypothesis, the analysis of membrane proteins from stationary-phase cells showed that a strain carrying this gene on a multicopy vector overproduces a protein of 30 kDa. This protein was recognized by antibodies directed against the N-terminal portion of the gene product. Considering its induction in response to heat shock and the apparent molecular weight of its product, this gene was designated HSP30.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Journal of cancer research and clinical oncology 119 (1993), S. 675-684 
    ISSN: 1432-1335
    Keywords: Xeroderma pigmentosum fibroblasts ; DNA repair ; Recombinant DNA ; cDNA libraries ; Phage λgt10 ; pBluescript vector in vitro transcripts ; Differential hybridization ; Mitochondrial neuromyopathies ; Genetic defects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract cDNA libraries constructed from cytoplasmic RNA of normal and xeroderma pigmentosum (XP) fibroblast strains were screened for differential gene expression. XP fibroblast strains included one representative of the complementation groups A, C, D, and one XP variant strain. The XP λgt10 cDNA libraries were differentially screened with in vitro transcripts made from cDNA in the pBluescript vector using both the same XP strain and the normal fibroblast strain. Eight differential clones were detected in the libraries of the XP group A, D, and C strains, which caused stronger signals when probed with transcripts from XP strains than with those from the normal strain. The cDNA clones were sequenced. Seven of the eight clones detected coded for three mitochondrial genes: subunit I of cytochromec oxidase (complex IV of the respiratory chain), apocytochromeb (subunit of complex III), and 16-S rRNA. Two clones representing essentially (a) subunit I of cytochromec oxidase and (b) 16-S rRNA diverged from the sequence of the human mitochondrial genome present in the data-base libraries. Clone a exhibited a transition mutation, clone b reflected a transcript of a mitochondrial genome rearranged in the 16-S rRNA gene, including four nucleotides of the adjacent tRNALeu gene. The apparently enhanced expression of mitochondrial genes in XP cells, together with the changes in DNA sequence, seem to indicate that functions of the ATP-generating system were impaired. This defect may have originated from mutations due to lack of DNA repair. The data can be interpreted in the light of mitochondrial changes that cause human neuromyopathies to occur. In analogy to these diseases the neurological symptoms in XP might be explained by abnormal mitochondria.
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  • 13
    ISSN: 1573-5087
    Keywords: auxin ; Catharanthus roseus ; 2,4-dichlorophenoxyacetic acid (2,4-D) ; indole alkaloid ; protein synthesis ; two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstracts Total and neosynthesized proteins of periwinkle cell suspensions (Catharanthus roseus) were first investigated in cells grown in a 2,4-D-containing medium. Analysis of total (silver-stained) proteins by two-dimensional gel electrophoresis revealed that the levels of seventeen polypeptides were altered during the growth cycle of the cells. Analysis of in vivo [35S]-methionine labeled polypeptides revealed differences in the synthesis of at least 35 polypeptides. Three polypeptides with molecular masses of 30, 35 and 39 kDa appeared to be specific markers of the early stationary phase. In a second sequence of experiments, cells were grown in a 2,4-D-free medium. Alterations in protein synthesis were observed: several polypeptides were expressed earlier in the 2,4-D-starved cells than in control cells; the synthesis of at least two specific polypeptides was increased in cells grown in 2,4-D-free medium, whereas the synthesis of three other polypeptides (molecular masses 33, 34 and 52.5 kDa) was switched on in these cells. As previous studies showed that 2,4-D depletion increased the alkaloid production in C. roseus cells, the present results may suggest that these polypeptides are implicated in the regulation of the alkaloid pathway.
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  • 14
    Electronic Resource
    Electronic Resource
    Springer
    Biodegradation 4 (1993), S. 231-240 
    ISSN: 1572-9729
    Keywords: bioremediation ; sediments ; Aroclor ; anaerobic microorganisms ; anaerobiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Reductive dechlorination is an advantageous process to microorganisms under anaerobic conditions because it is an electron sink, thereby allowing reoxidation of metabolic intermediates. In some organisms this has been demonstrated to support growth. Many chlorinated compounds have now been shown to be reductively dechlorinated under anaerobic conditions, including many of the congeners in commercial PCB mixtures. Anaerobic microbial communities in sediments dechlorinate Aroclor at rates of 3 µg Cl/g sediment × week. PCB dechlorination occurs at 12° C, a temperature relevant for remediation at temperate sites, and at concentrations of 100 to 1000 ppm. The positions dechlorinated are usually meta 〉 para 〉 ortho. The biphenyl rings, and the mono-ortho- and diorthochlorobiphenyls were not degraded after a one year incubation. Hence subsequent aerobic treatment may be necessary to meet regulatory standards. Reductive dechlorination of Arochlors does reduce their dioxin-like toxicity as measured by bioassay and by analysis of the co-planar congeners. The most important limitation to using PCB dechlorination as a remediation technology is the slower than desired dechlorination rates and no means yet discovered to substantially enhance these rates. Long term enrichments using PCBs as the only electron acceptor resulted in an initial enhancement in dechlorination rate. This rate was sustained but did not increase in serial transfers. Bioremediation of soil contaminated with Aroclor 1254 from a transformer spill was dechlorinated by greater than 50% following mixing of the soil with dechlorinating organisms and river sediment. It is now reasonable to field test reductive dechlorination of PCBs in cases where the PCB concentration is in the range where regulatory standards may be directly achieved by dechlorination, where a subsequent aerobic treatment is feasible, where any co-contaminants do not pose an inhibitory problem, and where anaerobic conditions can be established.
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  • 15
    ISSN: 1572-9729
    Keywords: aromatic hydrocarbons ; biodegradation ; bioremediation ; denitrification ; groundwater ; Pseudomonas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We characterized bacteria from contaminated aquifers for their ability to utilize aromatic hydrocarbons under hypoxic (oxygen-limiting) conditions (initial dissolved oxygen concentration about 2 mg/l) with nitrate as an alternate electron acceptor. This is relevant to current intense efforts to establish favorable conditions forin situ bioremediation. Using samples of granular activated carbon slurries from an operating groundwater treatment system, we isolated bacteria that are able to use benzene, toluene, ethylbenzene, orp-xylene as their sole source of carbon under aerobic or hypoxic-denitrifying conditions. Direct isolation on solid medium incubated aerobically or hypoxically with the substrate supplied as vapor yielded 103 to 105 bacteria ml−1 of slurry supernatant, with numbers varying little with respect to isolation substrate or conditions. More than sixty bacterial isolates that varied in colony morphology were purified and characterized according to substrate utilization profiles and growth condition (i.e., aerobic vs. hypoxic) specificity. Strains with distinct characteristics were obtained using benzene compared with those isolated on toluene or ethylbenzene. In general, isolates obtained from direct selection on benzene minimal medium grew well under aerobic conditions but poorly under hypoxic conditions, whereas many ethylbenzene isolates grew well under both incubation conditions. We conclude that the conditions of isolation, rather than the substrate used, will influence the apparent characteristic substrate utilization range of the isolates obtained. Also, using an enrichment culture technique, we isolated a strain ofPseudomonas fluorescens, designated CFS215, which exhibited nitrate dependent degradation of aromatic hydrocarbons under hypoxic conditions.
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  • 16
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 41 (1993), S. 625-632 
    ISSN: 0006-3592
    Keywords: bioremediation ; biodegradation ; soil ; sorption/desorption ; intraparticle diffusion ; pollution ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: To determine when intraparticle diffusion and sorption can influence the rate of biodegradation, we consider the biodegradation of a pollutant diffusing into or out of porous aggregates suspended in a liquid medium, where the reactant is metabolized by bacteria. The pollutant that diffuses into the aggregates obeys a sorption-desorption equilibrium isotherm at sites on inner pore surfaces. The governing partial differential equations for the transient process describe (a) the local equilibrium sorption-desorption and the diffusion of the pollutant in the porous aggregate, (b) the mass transfer of the pollutant from the external surface of the spherical aggregates to the reaction medium, and (c) the biodegradation of the pollutant in the external medium. Illustrative calculations are presented for a linear sorption calculations are presented for a linear sorption isotherm and first-order biodegradation kinetics. A dimensionless group, comprised of the diffusion coefficient, biodegradation rate coefficient, aggregate characteristics length (radius), and adsorption capacity, serves as a criterion for determining when intraparticle diffusion can be ignored. The model provides a realistic description of experimental data for biodegradation of a pollutant subject to intraparticle diffusion and sorption. © 1993 John Wiley & Sons, Inc.
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  • 17
    Electronic Resource
    Electronic Resource
    Springer
    Biodegradation 3 (1992), S. 161-170 
    ISSN: 1572-9729
    Keywords: bioremediation ; cadmium ; heavy metals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Cadmium pollution arises mainly from contamination of minerals used in agriculture and from industrial processes. The usual situation is of large volumes of soil and water that are contaminated with low — but significant — concentrations of cadmium. Therefore, detoxification of the polluted water and soil involves the concentration of the metal, or binding it in a way that makes it biologically inert. Cadmium is one of the more toxic metals, that is also carcinogenic and teratogenic. Its effects are short term, even acute (diseases like Itai-itai), or long term. The long term effects are intensified due to the fact that cadmium accumulates in the body. This paper describes a study involving several hundred cadmium-resistant bacterial isolates. These bacteria could be divided into three groups—the largest group consisted of bacteria resistant to cadmium by effluxing it from the cells. The bacteria of the other two groups were capable of binding cadmium or of detoxifying it. We concentrated on one strain that could bind cadmium very efficiently, depending on the bacterial biomass and on the pH. This strain could effectively remove cadmium from contaminated water and soil.
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  • 18
    ISSN: 1572-9729
    Keywords: petroleum ; bioremediation ; hydrocarbon pollution ; bioemulsifiers ; petroleum microbiology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Microbial degradation of hydrocarbons is a multiphase reaction, involving oxygen gas, water-insoluble hydrocarbons, water, dissolved salts and microorganisms. The fact that the first step in hydrocarbon catabolism involves a membrane-bound oxygenase makes it essential for microorganisms to come into direct contact with the hydrocarbon substrate. Growth then proceeds on the hydrocarbon/water interface. Bacteria have developed two general strategies for enhancing contact with water-insoluble hydrocarbons: specific adhesion mechanisms and production of extracellular emulsifying agents. Since petroleum is a complex mixture of many different classes of hydrocarbons, of which any particular microorganism has the potential to degrade only part, it follows that the microorganisms must also have a mechanism for desorbing from used' oil droplets. The major limitations in bioremediation of hydrocarbon-contaminated water and soil is available sources of nitrogen and phosphorus. The usual sources of these materials, e.g. ammonium sulfate and phosphate salts, have a high water solubility which reduces their effectiveness in open systems because of rapid dilution. We have attempted to overcome this problem by the use of a new controlled-release, hydrophobic fertilizer, F-1, which is a modified urea-formaldehyde polymer containing 18% N and 10% P as P2O5. Microorganisms were obtained by enrichment culture that could grow on crude oil as the carbon and energy source and F-1 as the nitrogen and phosphorus source. The microorganisms and the F-1 adhered to the oil/water interface, as observed microscopically and by the fact that degradation proceeded even when the water phase was removed and replaced seven times with unsupplemented water — a simulated open system. Strains which can use F-1 contain a cell-bound, inducible enzyme which depolymerizes F-1. After optimizing conditions in the laboratory for the use of F-1 and the selected bacteria for degrading crude oil, a field trial was performed on an oil contaminated sandy beach between Haifa and Acre, Israel, in the summer of 1992. The sand was treated with 5 g F-1 per kg sand and inoculated with the selected bacteria; the plot was watered with sea water and plowed daily. After 28 days the average hydrocarbon content of the sand decreased from 5.1 mg per g sand to 0.6 mg per g sand. Overall, there was an approx. 86% degradation of pentane extractables as demonstrated by dry weight, I.R. and GLC analyses. An untreated control plot showed only a 15% decrease in hydrocarbons. During the winter of 1992, the entire beach (approx. 200 tons of crude oil) was cleaned using the F-1 bacteria technology. The rate of degradation was 0.06 mg g-1 sand day-1 (10°C) compared to 0.13 mg g-1 sand day-1 during the summer (25°C).
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  • 19
    ISSN: 1432-0584
    Keywords: β-globin ; Unstable hemoglobins ; Polymerase chain reaction ; DGGE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Characterization of unstable hemoglobins by protein analysis is often difficult. However, it is facilitated by DNA analysis, especially in the case of hyperunstable β-chain variants, which produce a β-thalassemia phenotype. We have applied an efficient strategy to the detection of such variants at the DNA level, based on computer-designed denaturing gradient gel electrophoresis (DGGE) of amplified DNA fragments. This approach makes it possible to detect any anomaly in the β-globin gene. We describe the use of the DGGE method for rapid characterization of β-chain variants and report a new missense mutation in the β-globin gene third exon, β 127 CAG-CGG/Gln-Arg, which is responsible for the synthesis of a highly unstable hemoglobin.
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  • 20
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 84 (1992), S. 573-578 
    ISSN: 1432-2242
    Keywords: Common wheat ; RAPD ; DGGE ; DNA polymorphism ; DNA markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A DNA marker detection strategy that allows the rapid, efficient resolution of high levels of polymorphism among closely related lines of common wheat (Triticum aestivum) has been developed to circumvent the apparent lack of restriction fragment length polymorphism in many important self-pollinated crop species. The technique of randomly amplified polymorphic DNA (RAPD) was combined with a denaturing gradient gel electrophoresis system (DGGE) to explore DNA sequence polymorphisms among different genotypes of wheat. Of the 65 primer combinations used for the polymerase chain reaction (PCR) amplifications, over 38% of them produced readily detectable and reproducible DNA polymorphisms between a spring wheat line, SO852, and a winter wheat variety, ‘Clark’. A high level of polymorphism was observed among a number of commercial varieties and breeding lines of wheat. This procedure was also used to detect polymorphisms in a recombinant inbred population to test the feasibility of its application in genome mapping. This DNA polymorphism detection system provides an opportunity for pedigree analysis and fingerprinting of developed wheat lines as well as construction of a high density genetic map of wheat. Without the need for 32P and sophisticated DNA extraction procedures, this approach should make it feasible to utilize marker-based selection in a plant breeding program.
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  • 21
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Shotgun cloning ; Filamentous fungi ; Promoter sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A transformation system for the thermophilic cellulolytic fungus Talaromyces sp. CL240 has been developed, using the phleomycin resistance gene from Streptoalloteichus hindustanus (Sh ble) as a dominant selectable marker. The plasmids (pAN8-1 and pUT720) carrying the Sh ble gene under the control of the Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter, allowed selection of phleomycin-resistant transformants. A new promoter sequence cloned from chromosomal DNA of Trichoderma reesei (pUT737) was also able to drive efficient expression of the Sh ble gene in Talaromyces sp. CL240, resulting in the selection of transformants that were highly resistant to phleomycin.
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  • 22
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    Molecular genetics and genomics 232 (1992), S. 498-504 
    ISSN: 1617-4623
    Keywords: luminescence ; Recombinant DNA ; Grampositive organisms ; Shuttle vectors ; luc and lux genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A group of vectors for luciferase expression in Bacillus subtilis was constructed. So far, only bacterial luciferases have been expressed in Bacillus, but in this study we wanted also to express genes encoding eukaryotic luciferases to perform direct comparisons of the light levels produced by the two different systems in B. subtilis. The vectors constructed can replicate both in Escherichia coli and B. subtilis, and the luciferase expression is strictly regulated due to the dual plasmid system used. Nearly a 100-fold increase in light production compared to previous results was achieved when genes encoding bacterial luciferase were inserted into the constructs and transformed into B. subtilis. An additional tenfold increase in light production was obtained when luciferase genes from the North American firefly (Photinus pyralis) or a click beetle (Pyrophorus plagiophtalamus) were introduced in a similar fashion into B. subtilis. Measurement of the light emission was performed without disruption of bacterial cells in a real-time manner, which is a common feature when working with all of these constructions. Structures of the shuttle vector constructs and results from light emission measurements are presented.
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  • 23
    ISSN: 1617-4623
    Keywords: Secretion ; Recombinant DNA ; Hemolysin ; HlyB/H1yD complementation ; OmpT protease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A fusion gene (ces-hlyA s) was constructed by ligating the genetic information for the C-terminal 60 amino acids (hlyA s) ofEscherichia coli hemolysin (H1yA) to the ces gene for a cholesterol esterase/lipase (CE) from aPseudomonas species. Part (about 30 %) of the expressed fusion protein CE-H1yAs was secreted inE. coli carryinghlyB andhlyD genes. Following the insertion between the reporter gene andhlyA s of a linker sequence that contains the information for potential cleavage sites for the outer membrane protease OmpT, two different fusion proteins (PhoA-H1yAs and CE-HlyAs) were shown to be cleaved by OmpT between the two parts during H1yB/H1yD-mediated secretion. Processed PhoA and CE accumulated in the supernatant. The efficiency of cleavage by OmpT was considerably improved by increasedompT gene dose. It was further shown that OmpT preferentially recognizes potential cleavage sites within the linker sequence.
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  • 24
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    Journal of aquatic ecosystem stress and recovery 1 (1992), S. 253-262 
    ISSN: 1573-5141
    Keywords: bacteria ; factor analysis ; path analysis ; aquatic ; gene probe ; bioremediation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Studies on aquatic ecosystems at the trophic level of bacteria include population succession and the movement of species through the water column. Factor and path analysis of environmental parameters and the bacterial profiles indicate that the bacterial populations are under the control of environmental factors. The most important environmental factors in the Canadian study reported here are temperature followed by oxygen levels, nutrient levels, and ion concentrations. A major revolution in investigative approaches has begun in aquatic bacterial population studies using technology based on molecular methods. Finger print analysis of bacterial 16S RNA (molecular phylogeny) has not only changed the classification of bacteria but also the approach to solving environmental problems. The bacterial groups have been placed into species that are more functionally and ecologically aligned. Uncultured mixed biomass can be examined by gene probes for both procaryotes and eucaryotes to identify specific nucleotide sequences. Aquatic ecosystem health is maintained by the balanced biota and the process of biodegradation is an important stage in bioremediation. Control of toxic wasters in the waters and groundwaters can be accomplished byin situ bioremediation using indigenous microorganisms as demonstrated by the field study reported by Litchfieldet al. (1990).
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  • 25
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    Journal of industrial microbiology and biotechnology 10 (1992), S. 185-190 
    ISSN: 1476-5535
    Keywords: β-Glucosidase ; C. flavigena ; Cellobiase ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Plasmid-coded β-glucosidase produced byEscherichia coli was characterized and compared to the enzyme produced byCellulomonas flavigena. Cell-free extracts, non-denaturing PAGE and 5-bromo-4-chloro-3-indolyl-β-d-glucopyranoside (X-glu) as substrate were used to compare both enzymes. The β-glucosidase was assayed for cellobiose andp-nitrophenyl-glucopyranoside (PNPG). Cellobiose hydrolysis was performed at 50°C for the enzyme fromC. flavigena and at 37°C for that fromE. coli pJS3, both with an optimal pH of 6.5. For PNPG hydrolysis, the optimal conditions were pH 5.5 and 37°C for both cell extracts. Most of the β-glucosidase activity was intracellular. When cultures ofC. flavigena were grown with cellobiose or carboxymethylcellulose (CMC) as inducers, the expression of β-glucosidase was increased considerably.E. coli pJS3 produces a cellobiase which hydrolyzes cellobiose and PNPG. TheK m values for cellobiose and PNPG indicated that the β-glucosidase activity ofC. flavigena had a higher affinity for cellobiose as substrate, whereas the β-glucosidase fromE. coli pJS3 showed higher affinity for PNPG.
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  • 26
    ISSN: 1572-9729
    Keywords: bioremediation ; inorganic mercury ; organomercury
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Bacterially mediated ionic mercury reduction to volatile Hg0 was shown to play an important role in the geochemical cycling of mercury in a contaminated freshwater pond. This process, and the degradation of methylmercury, could be stimulated to reduce the concentration of methylmercury that is available for accumulation by biota. A study testing the utility of this approach is described.
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  • 27
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    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 31 (1992), S. 161-169 
    ISSN: 1040-452X
    Keywords: Sperm cell ; Recombinant DNA ; Fertilization ; Genetic transformation ; Transgenic animals ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Epididymal sperm cells, incubated with plasmid DNA, showed a spontaneous tendency to interact with the exogenous nucleic acid. We have investigated the molecular basis of such interaction. Exogenous DNA is taken up by sperm cells over a 15- to 20-min period and is specifically localized on the nuclear area of the sperm head. DNA was reversibly bound to spermatozoa since it can be competed out by excess of cold competitor DNA or by other polyanions as heparin and dextran sulphate. By contrast, poly-L-lysine, a polycation, favours the uptake. DNA molecules of large size (7 kb) were preferentially taken up as compared to smaller ones (150-750 bp). Acidic proteins were also taken up and concentrated, as for DNA, at the nuclear level. These data strongly suggested that ionic interactions may occur between foreign molecules and a substrate located in the sperm head. On the basis of Southwestern analysis, a sperm head protein(s) of 30-35 KD is identified as potential substrate for exogenous DNA binding. Moreover, we have found that seminal plasma contains factor(s) which abolish sperm permeability, exerting a powerful inhibitor effect on DNA uptake. The presence of a specific binding protein for the DNA and of a factor inhibiting such interaction support the existence of a mechanism controlling, through specific factors, the sperm-DNA interaction.
    Additional Material: 9 Ill.
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  • 28
    ISSN: 1572-9729
    Keywords: hydrocarbon biodegradation ; bioassay ; radiorespirometry ; bioremediation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Following the EXXOn Valdez oil spill, a radiorespirometric protocol was developed at the University of Alaska Fairbanks (UAF) to assess the potential for microorganisms in coastal waters and sediments to degrade hydrocarbons. The use of bioremediation to assist in oil spill cleanup operations required microbial bioassays to establish that addition of nitrogen and phosphorus would enhance biodegradation. A technique assessing 1-14C-n-hexadecane mineralization in seawater or nutrient rich sediment suspensions was used for both of these measurements. Hydrocarbon-degradation potentials were determined by measuring mineralization associated with sediment microorganisms in sediment suspended in sterilized seawater and/or marine Bushnell-Haas broth. Production of 14CO2 and CO2 was easily detectable during the first 48 hours with added hexadecane levels ranging from 10 to 500 mg/l of suspension and dependent on the biomass of hydrocarbon degraders, the hydrocarbon-oxidation potential of the biomass and nutrient availability. In addition to assessment of the hydrocarbon-degrading potential of environmental samples, the radiorespirometric procedure, and concomitant measurement of microbial biomass, has utility as an indicator of hydrocarbon contamination of soils, aqueous sediments and water, and can also be used to evaluate the effectiveness of bioremediation treatments.
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  • 29
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    European archives of psychiatry and clinical neuroscience 240 (1991), S. 197-203 
    ISSN: 1433-8491
    Keywords: Schizophrenia ; Recombinant DNA ; Genetic markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Interest in genetic marker studies of schizophrenia has been considerably enhanced by the advent of recombinant DNA technology, which has dramatically increased the number of available markers. In the present paper, we review studies that have been carried out using classical markers as well as the more recent molecular studies. The problems that arise when schizophrenia is studied in this way are discussed and attempts are made to account for some of the conflicting findings in this area.
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  • 30
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Gene cloning ; Ubiquitin fusion gene ; Ubiquitin extension protein ; Expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The genome of Tetrahymena pyriformis has been shown to contain a ubiquitin multigene family consisting of several polyubiquitin genes and at least one ubiquitin fusion gene. We report here the isolation and characterization of one genomic clone (pTUl1), that encodes a ubiquitin extension protein. A comparison of the predicted amino acid sequence of the ubiquitin extension protein gene of T. pyriformis with those from other organisms indicated a high degree of homology. However, the Tetrahymena ubiquitin extension protein contains 53 and not 52 amino acids. This feature is different from all ubiquitin 52-amino-acid extension protein genes thus far sequenced. Furthermore, we found an array of four cysteine residues similar to those found in nucleic acid binding proteins. Also, the C-terminal sequence possesses a conserved motif which may represent a nuclear translocation signal. The ubiquitin 53-amino-acid extension protein gene encodes the smallest class of ubiquitin mRNAs in T. pyriformis.
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  • 31
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    Molecular genetics and genomics 227 (1991), S. 28-32 
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Antibiotic production ; Genetic organization ; Directed mutant screen ; Prodigiosin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Fifteen mutants ofStreptomyces coelicolor A3(2) blocked in both the bipyrrole branch (redA) and a second site specific to the undecylprodigiosin pathway were characterized. Some of the mutants were ordered biosynthetically based on cosynthesis experiments. Complementation of each of the mutants with wild-type DNA cloned in low- and high-copy number plasmid vectors allowed the mutants to be separated into 12 new classes which are physically clustered within approximately 37 kb on theS. coelicolor genome. Early-step biosynthetic genes are centrally located and are flanked by later-step and regulatory genes.
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  • 32
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    New forests 5 (1991), S. 359-367 
    ISSN: 1573-5095
    Keywords: indole-3-butyric acid (IBA) ; 2,4-dichlorophenoxyacetic acid (2,4-D) ; bareroot seedlings ; root development ; root dip ; Pinus ponderosa (Dougl. ex Laws.) ; Celtis occidentalis (L.) ; hygroscopic gel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Greenhouse and field studies were conducted to determine the effects of indole3-butyric acid (IBA) and 2,4-dichlorophenoxyacetic acid (2,4-D) on root development and survival of 3+0 bareroot Pinus ponderosa (Dougl. ex Laws.) and 2+0 bareroot Celtis occidentalis (L) seedlings. In the greenhouse, 100 and 200 ppm. IBA and 10 and 50 ppm 2,4-D were applied to seedlings through a root dip in an auxin hygroscopic gel mix. A randomized complete block analysis of variance indicated that IBA gel treatments increased root volume and root dry weight in hackberry and had no effect on ponderosa pine; 2,4-D gel treatments had detrimental effects on both species. In the field, IBA gel treated ponderosa pine seedlings exhibited improved height and survival relative to the non-treated control seedlings. There was no detectable improvement in survival in the field for hackberry. However, diameter and stem dry weight were lower for hackberry seedlings treated with the gel dip alone.
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  • 33
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    European journal of epidemiology 7 (1991), S. 213-221 
    ISSN: 1573-7284
    Keywords: Rickettsiae ; Rickettsial genetics ; Genes ; Recombinant DNA ; Cloning ; Electroporation ; Transformation Rickettsia ; Coxiella ; Rochalimaea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Classical genetic approaches useful with free-living bacteria are difficult to apply to the rickettsiae. Although rickettsial mutants have been isolated over the years, the genetic basis of these mutants is unknown, limiting their usefulness. The application of molecular biological techniques to rickettsial studies has provided the opportunity to isolate and study specific genes. Genes encoding metabolic enzymes from rickettsiae were cloned in Escherichia coli and shown to retain their regulatory properties, suggesting that recombinant DNA technology may be useful for studies of rickettsial enzymes and regulatory mechanisms. The potential use of rickettsial surface components, or virulence factors as possible antigens for protective subunit vaccines, has led to the cloning and expression in E. coli, of rickettsial chromosomal and plasmid genes encoding outer membrane proteins. The number of genes characterized in recent years has increased dramatically giving rise to an increasing source of information on rickettsial gene structure. Plasmids have only been identified in C. burnetii and possibly Rochalimaea quintana. The plasmid sequences present in C. burnetii are highly conserved suggesting that they are important to the growth and virulence of this organism. To understand the role of genes in the rickettsia-host relationship, it is critical that a genetic exchange system be developed. The recent description of transformation of R. quintana by electroporation is an important first step in this direction. The ability to introduce genetic material is necessary to address questions that cannot be resolved by studying rickettsial gene expression in E. coli.
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  • 34
    ISSN: 1476-5535
    Keywords: Fermentation ; Recombinant DNA ; Phage λp L promoter ; Expression vector ; α1-Antitrypsin ; Malaria vaccine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The major leftward early promoter of phage λp L, has frequently been used to drive expression of heterologous genes inEscherichia coli.p L is typically maintained fully repressed by the lambda cl protein. When induction of heterologous protein synthesis is desired, one of several potential mechanisms of destroying cl function is employed and the expression of the foreign gene commences. One method of derepressingp L involves exposing cells to nalidixic acid, which results in the “activation” of RecA protein and the subsequent RecA-mediated proteolytic cleavage of cl. Activated RecA also mediates the cleavage of theE. coli LexA protein, resulting in induction of the SOS regulon (at least 15E. coli genes, includingrec A). We have examined the effect of two chromosomal mutations on the productivity of nalidixic acid inductions. One of the tested mutations (recA o) increased the intracellular concentration of RecA prior to induction; the other (lexAind−) resulted in a mutated lexA protein insensitive to RecA-mediated cleavage. These mutations were introduced into a strain carrying acl+ defective lysogen. Synthesis of two heterologous proteins, human α1-antitrypsin and a fusion protein partially derived from thePlasmodium falciparum circumsporozooite surface antigen, was examined in the wild-type and mutant strains. The maximum α-1 antitrypsin concentration achieved was improved by 50% when therecA o strain was used rather than the wild type; however; only smaller changes (20% or less) in the maximum concentration of the malaria fusion protein wer observed. Use of thelexAind− strain resulted in a decrease in the maximum concentration attained for both heterologous products.
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  • 35
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    Current genetics 17 (1990), S. 223-227 
    ISSN: 1432-0983
    Keywords: Orotidine-5′-phosphate decarboxylase ; Cephalosporium acremonium ; Recombinant DNA ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned the Cephalosporium acremonium pyr4 gene by cross-hybridization with the equivalent gene from Neurospora crassa, the closest relative from which this gene is available. The C. acremonium pyr4 gene complements an E. coli pyrF mutant lacking orotidine-5′-phosphate decarboxylase (OMPdecase), and most probably does not contain introns. Maxicell analysis in E. coli shows that it encodes a 46 kDa polypeptide. The C. acremonium OMPdecase contains a highly conserved pentadecapeptide characteristic for this category of enzyme. Extensive sequence comparison suggests an important role of this region in enzymatic activity.
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  • 36
    ISSN: 1617-4623
    Keywords: Monooxygenase ; Recombinant DNA ; Overexpression ; Upstream reading frames ; Complementation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The gene coding for benzoate-para-hydroxylase (bphA) of Aspergillus niger was cloned using differential hybridisation techniques and complementation of mutants deficient in this enzyme activity. The nucleotide sequence of the gene was determined, the presence of two introns was shown and the transcription start and termination sites were determined. The structure of the mRNA upstream from the long open reading frame (ORF) is unusual. It contains two small, overlapping ORFs whose function is unknown. Comparison of the deduced amino acid sequence of the protein with the sequences present in the databanks, indicated a significant similarity of BPH to the superfamily of cytochrome P450 enzymes. Further analysis revealed that this protein is a member of a new P450 gene family designated P450LIII. The gene is designated CYP53. To increase the BPH activity of A. niger, multiple copies of the bphA gene were introduced into the genome of a recipient strain by transformation. Although increased intracellular levels of the BPH protein could be detected, the BPH enzyme activity was decreased, suggesting titration of another essential component.
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  • 37
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    Molecular genetics and genomics 222 (1990), S. 249-256 
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Plant virus ; T7 promoter ; In vitro transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Full-length cDNA copies of cucumber mosaic virus (CMV) RNAs 1 and 2 of the Fny strain were constructed from partial cDNA clones and were cloned downstream of bacteriophage T7 promoters. In one pair of clones, transcription proceeded from an unaltered T7 promoter such that in vitro transcripts representing RNAs 1 and 2 contained an additional 17 nucleotides at their 5′ termini. In a second pair of clones, the T7 promoter/cDNA junction was altered by oligonucleotide-directed mutagenesis such that the in vitro transcripts contained only an additional G residue at their 5′ ends. In addition, a full-length cDNA copy of Fny-CMV RNA 3 was constructed from two overlapping cDNA clones and was cloned downstream of an altered T7 promoter such that the resultant in vitro transcripts also contained only an additional G residue at their 5′ ends. In vitro transcripts derived from all clones contained an additional C residue at their 3′ ends. In vitro transcripts representing RNAs 1, 2 and 3 which contained an additional residue at each terminus were shown to be infectious together in several hosts of CMV.
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  • 38
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Vaccine development ; Carrier protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Hypervariable regions (HRs) of the major subunit of F11 fimbriae were exploited for insertion of foreign epitopes. Two insertion vectors were created that contain a unique cloning site in HR1 or HR4 respectively. Several oligonucleotides, coding for antigenic determinants derived from different pathogens, were cloned in both insertion vectors. Hybrid fimbrial subunits were generally shown to be assembled in fimbriae when the length of the inserted peptide did not exceed 14 amino acids. The inserted peptides appeared to be exposed in the fimbrial filament. One hybrid fimbrial protein induced detectable levels of antibodies against the inserted epitope if injected into mice.
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  • 39
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    Biodegradation 1 (1990), S. 283-290 
    ISSN: 1572-9729
    Keywords: bioremediation ; metabolites ; microbial degradation ; naphthalene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A Mycobacterium sp. isolated from oil-contaminated sediments was previously shown to mineralize 55% of the added naphthalene to carbon dioxide after 7 days of incubation. In this paper, we report the initial steps of the degradation of naphthalene by a Mycobacterium sp. as determined by isolation of metabolites and incorporation of oxygen from 18O2 into the metabolites. The results indicate that naphthalene is initially converted to cis- and trans-1,2-dihydroxy-1,2-dihydronaphthalene by dioxygenase and monooxygenase catalyzed reactions, respectively. The ratio of the cis to trans-naphthalene dihydrodiol isomers was approximately 25:1. Thin layer and high pressure liquid chromatographic and mass spectrometric techniques indicated that besides the cis- and trans-1,2-dihydroxy-1,2-dihydronaphthalene, minor amounts of ring cleavage products salicylate and catechol were also formed. Thus the formation of both cis and trans-naphthalene dihydrodiols by the Mycobacterium sp. is unique. The down-stream reactions to ring cleavage products proceed through analogous dioxygenase reactions previously reported for the bacterial degradation of naphthalene.
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