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At least 163 records · Page 9Linked to original sources
PubMed1999

Production of new/modified proteins in transgenic plants.

In recent years, plant biotechnology has almost reached maturity. Transgenic plants engineered to be herbicide- or insect-resistant are outcompeting conventional crop plants and pest managing strategies leading to a major rethinking of the chemical industry. Due to worldwide efforts to study genome function, almost any gene of interest is, or will soon be available. Thus, identification of gene function will be the major challenge of the next few years. In combination with established gene-delivery systems and desired promoter and targetting sequences, gene discovery will open a fascinating and new field of crop plant design. Transgenic plants engineered to produce superior polypeptides have already been created and the first examples are entering clinical and industrial trials.

Amino Acids↗
PubMed1998

Expression and immunolocalisation of the snowdrop lectin, GNA in transgenic rice plants.

Transgenic rice (Oryza sativa L.) plants generated through particle bombardment expressed high levels of an insecticidal protein (the snowdrop lectin, GNA) directed against sap-sucking insects. Engineered plants expressed GNA either constitutively or in a tissue specific manner, depending on the nature of the promoter used to drive expression of the gene. We used specific antibodies raised against GNA to localize its expression in phloem tissue in plants engineered with the rice sucrose synthase promoter driving GNA expression. We report here molecular, biochemical and immunological analyses for fifteen independently-derived transformants out of more than 200 plants we generated.

Gene Expression↗
PubMed2003

Functional analysis of cystathionine gamma-synthase in genetically engineered potato plants.

In plants, metabolic pathways leading to methionine (Met) and threonine diverge at the level of their common substrate, O-phosphohomoserine (OPHS). To investigate the regulation of this branch point, we engineered transgenic potato (Solanum tuberosum) plants affected in cystathionine gamma-synthase (CgS), the enzyme utilizing OPHS for the Met pathway. Plants overexpressing potato CgS exhibited either: (a) high transgene RNA levels and 2.7-fold elevated CgS activities but unchanged soluble Met levels, or (b) decreased transcript amounts and enzyme activities (down to 7% of wild-type levels). In leaf tissues, these cosuppression lines revealed a significant reduction of soluble Met and an accumulation of OPHS. Plants expressing CgS antisense constructs exhibited reductions in enzyme activity to as low as 19% of wild type. The metabolite contents of these lines were similar to those of the CgS cosuppression lines. Surprisingly, neither increased nor decreased CgS activity led to visible phenotypic alterations or significant changes in protein-bound Met levels in transgenic potato plants, indicating that metabolic flux to Met synthesis was not greatly affected. Furthermore, in vitro feeding experiments revealed that potato CgS is not subject to feedback regulation by Met, as reported for Arabidopsis. In conclusion, our results demonstrate that potato CgS catalyzes a near-equilibrium reaction and, more importantly, does not display features of a pathway-regulating enzyme. These results are inconsistent with the current hypothesis that CgS exerts major Met metabolic flux control in higher plants.

Carbon-Oxygen Lyases↗
PubMed2004

[Determination of fumes and their elements from flux cored arc welding].

BACKGROUND: The aim of this work was to assay the concentration levels and composition of welding fumes, released during flux cored arc welding, to assess exposure of welders. MATERIALS AND METHODS: Concentrations and welding fume components, such as iron, manganese, chromium (including the soluble and chromium VI), nickel, copper, calcium, aluminium, barium, and fluorides (including hydrogen fluoride) were determined in the air of six industrial plants (shipyards, mechanical engineering plants and a power station) at the breathing zones of the welders who used 10 types of wires during flux cored arc welding. The following determination methods were used: gravimetry (fumes), AAS (metals), and spectrophotometry (chromium VI, fluorides--including hydrogen fluoride). RESULTS: The results made it possible to determine the relationship between concentrations of welding fume and its elements, and to assess worker's exposure. Time weighted average concentrations of the welding fumes and its elements at the worker's breathing zone were: mg/m3: dust 0.2-24.3; Fe 0.2-6.7; Mn 0.01-1.8; Cr 0.004-0.5 (mainly Cr III); Ca 0.004-2.5; Ni < or = 0.004; Cu < 0.002-0.05; Al < 0.14-0.4; Ba < 0.14; F- 0.07-0.43. CONCLUSIONS: The welders using some types of flux cored welding wires worked in conditions harmful to their health owing to the considerably exceeded TLV value for fume and MAC values for manganese, and occasional slightly excessive MAC values for calcium and iron.

Air Pollutants, Occupational↗
PubMed1996

Assessment of the allergenic potential of foods derived from genetically engineered crop plants.

This article provides a science-based, decision tree approach to assess the allergenic concerns associated with the introduction of gene products into new plant varieties. The assessment focuses on the source from which the transferred gene was derived. Sources fall into three general categories: common allergenic food proteins; less common allergenic foods or other known allergen sources; and sources with no history of allergenicity. Information concerning the amino acid sequence identity to known allergenic proteins, in vitro and/or in vivo immunologic assays, and assessment of key physiochemical properties are included in reaching a recommendation on whether food derived from the genetically modified plant variety should be labeled as to the source of the transferred gene. In the end, a balanced judgement of all the available data generated during allergenicity assessment will assure the safety of foods derived from genetically engineered crops. Using the approaches described here, new plant varieties generated by genetic modification should be introduced into the marketplace with the same confidence that new plant varieties developed by traditional breeding have been introduced for decades.

Allergens↗
PubMed2005

Metabolic engineering of new fatty acids in plants.

Metabolic engineering of plants to express high levels of new fatty acids that are of nutritional and industrial importance has proven to be highly challenging. Significant advances have been made recently, however, particularly in the development of the first plant oils to contain long-chain polyunsaturated fatty acids, such as arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid. Methods of increasing the accumulation of Delta12-modified fatty acids synthesized by transgenically expressed FAD2-like enzymes have also been investigated. Biochemical analyses of plants that express these introduced fatty-acid metabolic pathways have highlighted the central importance of ensuring the removal of novel fatty acids from their site of synthesis on phosphatidylcholine to enable their further modification, exclusion from membrane lipids and accumulation in seed triacylglycerols.

Fatty Acids, Nonesterified↗
PubMed1990

Two outbreaks of Legionnaires' disease in Bolton Health District.

In 1988, there were two outbreaks of legionellosis in Bolton Health District. Altogether 37 cases of Legionnaires' disease and 23 cases of non-pneumonic legionellosis were identified. Twenty-five patients with Legionnaires' disease were associated with an engineering plant, 4 with Bolton town centre, and 8 with both the plant and town centre. Twenty-two people with non-pneumonic legionellosis were linked with the engineering plant and one with the plant and the town centre. A case-control study carried out among 37 employees with legionellosis and 109 control subjects at the plant showed that infection was associated with one of the 15 cooling towers on the site. Legionella pneumophila indistinguishable by serological and genetic typing methods was isolated from this cooling tower and from sputum samples from two patients. In the town centre, no one tower was linked with infection and L. pneumophila was not cultured from any of the nine towers identified. Control measures were implemented and to date there have been no further cases of legionellosis associated with Bolton Health District.

Adult↗
PubMed2001

Design of graph-based evolutionary algorithms: a case study for chemical process networks.

This paper describes the adaptation of evolutionary algorithms (EAs) to the structural optimization of chemical engineering plants, using rigorous process simulation combined with realistic costing procedures to calculate target function values. To represent chemical engineering plants, a network representation with typed vertices and variable structure will be introduced. For this representation, we introduce a technique on how to create problem specific search operators and apply them in stochastic optimization procedures. The applicability of the approach is demonstrated by a reference example. The design of the algorithms will be oriented at the systematic framework of metric-based evolutionary algorithms (MBEAs). MBEAs are a special class of evolutionary algorithms, fulfilling certain guidelines for the design of search operators, whose benefits have been proven in theory and practice. MBEAs rely upon a suitable definition of a metric on the search space. The definition of a metric for the graph representation will be one of the main issues discussed in this paper. Although this article deals with the problem domain of chemical plant optimization, the algorithmic design can be easily transferred to similar network optimization problems. A useful distance measure for variable dimensionality search spaces is suggested.

Algorithms↗
PubMed2006

Engineering plant-microbe symbiosis for rhizoremediation of heavy metals.

The use of plants for rehabilitation of heavy-metal-contaminated environments is an emerging area of interest because it provides an ecologically sound and safe method for restoration and remediation. Although a number of plant species are capable of hyperaccumulation of heavy metals, the technology is not applicable for remediating sites with multiple contaminants. A clever solution is to combine the advantages of microbe-plant symbiosis within the plant rhizosphere into an effective cleanup technology. We demonstrated that expression of a metal-binding peptide (EC20) in a rhizobacterium, Pseudomonas putida 06909, not only improved cadmium binding but also alleviated the cellular toxicity of cadmium. More importantly, inoculation of sunflower roots with the engineered rhizobacterium resulted in a marked decrease in cadmium phytotoxicity and a 40% increase in cadmium accumulation in the plant root. Owing to the significantly improved growth characteristics of both the rhizobacterium and plant, the use of EC20-expressing P. putida endowed with organic-degrading capabilities may be a promising strategy to remediate mixed organic-metal-contaminated sites.

Bacterial Proteins↗
PubMed2002

Metabolic engineering of plants: the role of membrane transport.

As plant cells are highly compartmentalized, the entrance and exit points of metabolic pathways frequently involve membrane passages of solutes. Transport proteins are often located in strategic positions to control whole pathways and have to be considered in the development of metabolic engineering strategies. Here, we discuss examples of pathways (in carbohydrate metabolism, amino acid and secondary compound synthesis, and mineral metabolism) in which membrane transport steps are considered to exert major control and in which transport proteins have been employed to manipulate metabolic fluxes.

Amino Acids↗
PubMed1996

Antibody production and engineering in plants.

Plants have always been important in the pharmaceutical industry, and plant biotechnology now offers new applications. The use of heterologous expression systems for production of recombinant proteins including antibodies is no longer novel. Attention has now turned towards the end-user, and specifically to engineering the antibody to precise requirements. In expanding the choice of production systems, plants offer unique alternatives for antibody users not only in the bulk production of antibody molecules but also in the ability to assemble full-length and complex, multimeric antibodies. Although the initial investment of time and effort is substantial compared to that of other expression systems, antibody production on an agricultural scale can be envisioned, and the economy of this approach opens many new areas for potential antibody applications in addition to their traditional research and pharmaceutical uses.

Animals↗
PubMed2005

Zinc finger nucleases: custom-designed molecular scissors for genome engineering of plant and mammalian cells.

Custom-designed zinc finger nucleases (ZFNs), proteins designed to cut at specific DNA sequences, are becoming powerful tools in gene targeting--the process of replacing a gene within a genome by homologous recombination (HR). ZFNs that combine the non-specific cleavage domain (N) of FokI endonuclease with zinc finger proteins (ZFPs) offer a general way to deliver a site-specific double-strand break (DSB) to the genome. The development of ZFN-mediated gene targeting provides molecular biologists with the ability to site-specifically and permanently modify plant and mammalian genomes including the human genome via homology-directed repair of a targeted genomic DSB. The creation of designer ZFNs that cleave DNA at a pre-determined site depends on the reliable creation of ZFPs that can specifically recognize the chosen target site within a genome. The (Cys2His2) ZFPs offer the best framework for developing custom ZFN molecules with new sequence-specificities. Here, we explore the different approaches for generating the desired custom ZFNs with high sequence-specificity and affinity. We also discuss the potential of ZFN-mediated gene targeting for 'directed mutagenesis' and targeted 'gene editing' of the plant and mammalian genome as well as the potential of ZFN-based strategies as a form of gene therapy for human therapeutics in the future.

Animals↗
PubMed2007

Methods for engineering resistance to plant viruses.

The development of genetically engineered resistance to plant viruses is a result of efforts to understand the plant-virus interactions involved in "crossprotection," a phenomenon observed with several plant virus diseases. Historically, expression of the coat protein gene of Tobacco mosaic virus in transgenic tobacco (Nicotiana tabacum) plants is the first example of transgene-mediated resistance to a plant virus. Subsequently, virus-derived sequences of several plant viruses were shown to confer virus resistance in experimental and/or natural hosts. For plant RNA viruses, virus complementary DNA sequences shown to confer resistance include wild-type genes, mutated genes that produced truncated protein products, and nontranslatable sense or antisense transcripts to various regions of the virus genome. Resistance also has been demonstrated for some viruses by mutant trans-dominant gene products, derived from the movement protein and replication-associated protein genes. In addition to virus-derived sequences, gene sequences of plant origin have also been used for transgenic resistance, and such resistance can be virus-specific, for instance, R genes isolated from resistant plant genotypes, or nonspecific, for example, ribosome inactivating proteins and proteinase inhibitors. Plantibodies and 2-5A synthetase, a class of proteins of mammalian origin, have also been useful in engineering plant virus resistance. In the case of transgenic resistance mediated by viral coat protein, the mechanism of resistance was suggested to operate during the early events of virus infection. However, transgene-mediated RNA silencing and generation of small interfering RNAs appears to be the primary mechanism that confers resistance to plant viruses. Despite the advantages of transgene-mediated resistance, current interest in the development and use of transgenic virus resistant plants is low in most parts of the world. However, because of its real potential, we believe that this technology will have more widespread and renewed interest in the near future.

Genetic Engineering↗
PubMed2005

Protocol: precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli.

Plant genome sequence data now provide opportunities to conduct molecular genetic studies at the level of the whole gene locus and above. Such studies will be greatly facilitated by adopting and developing further the new generation of genetic engineering tools, based on homologous recombination cloning in Escherichia coli, which are free from the constraints imposed by the availability of suitably positioned restriction sites. Here we describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs co-ordinately driven by locus-specific regulatory elements.

Journal Article↗