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PubMed1993

Genetic Variation within a Lotic Population of Janthinobacterium lividum.

An understanding of the genetic variation within and between populations should allow scientists to address many problems, including those associated with endangered species and the release of genetically modified organisms into the environment. With respect to microorganisms, the release of genetically engineered microorganisms is likely to increase dramatically given the current growth in the bioremediation industry. In this study, genetic variation within a lotic, bacterial population of Janthinobacterium lividum was measured with restriction fragment length polymorphism analysis. Chromosomal DNA from 10 Kettle Creek (Hawk Mountain Sanctuary, Kempton, Pa.) J. lividum isolates was digested with six restriction endonucleases and probed with a 7.5-kb pKK3535 fragment containing the E. coli rrnB rRNA operon. Genetic variation, as measured in terms of nucleotide diversity, was high within the population. The 0.0781 value for genetic variation was especially high given the conservative nature of the genetic probe. The average percent similarity among isolates within the population was 67.25%. Pairwise comparisons of nucleotide diversity values (pi) and similarity coefficients (F) yielded values ranging from 0.0032 to 0.1816 and 0.3363 to 0.9808, respectively. Putative clonemates were not present within the group of isolates; however, all isolates shared 14 fragments across a spectrum of six restriction enzymes. The presence of these common fragments indicates that restriction fragment length polymorphism analysis may provide population- or species-specific diagnostic markers for J. lividum. Data that suggest a plume effect with respect to the downstream movement of J. lividum are also presented. An increase in genetic variation within groups of isolates along the longitudinal gradient of Kettle Creek is also suggested.

Journal Article↗

PubMed2003

[Progress on biosafety assessment of marker genes in genetically modified foods].

Marker genes are useful in facilitating the detection of genetically modified organisms(GMO). These genes play an important role during the early identification stage of GMO development, but they exist in the mature genetically modified crops. So the safety assessment of these genes could not be neglected. In this paper, all the study on the biosafety assessment of marker genes were reviewed, their possible hazards and risks were appraised, and the marker genes proved safe were list too. GMO Labeling the is one important regulations for the development of genetically modified foods in the market. The accurate detecting techniques for GMO are the basis for setting up labeling regulation. In addition, some methods used to remove marker genes in genetically modified foods were introduced in the paper, which can eliminate their biosafety concern thoroughly.

Food, Genetically Modified↗

PubMed2005

Identification and quantification of three genetically modified insect resistant cotton lines using conventional and TaqMan real-time polymerase chain reaction methods.

As the genetically modified organisms (GMOs) labeling policies are issued in many countries, qualitative and quantitative polymerase chain reaction (PCR) techniques are increasingly used for the detection of genetically modified (GM) crops in foods. Qualitative PCR and TaqMan real-time quantitative PCR methods to detect and identify three varieties of insect resistant cotton, i.e., Mon531 cotton (Monsanto Co.) and GK19 and SGK321 cottons (Chinese Academy of Agricultural Sciences), which were approved for commercialization in China, were developed in this paper. Primer pairs specific to inserted DNAs, such as Cowpea trypsin inhibitor (CpTI) gene of SGK321 cotton and the specific junction DNA sequences containing partial Cry1A(c) gene and NOS terminator of Mon531, GK19, and SGK321 cotton varieties were designed to conduct the identified PCR assays. In conventional specific identified PCR assays, the limit of detection (LOD) was 0.05% for Mon531, GK19, or SGK321 in 100 ng of cotton genomic DNA for one reaction. Also, the multiplex PCR method for screening the three GM cottons was also established, which could save time and cost in practical detection. Furthermore, a real-time quantitative PCR assay based on TaqMan chemistry for detection of insect resistant gene, Cry1A(c), was developed. This assay also featured the use of a standard plasmid as a reference molecule, which contained both a specific region of the transgene Cry1A(c) and an endogenous stearoyl-acyl carrier protein desaturase (Sad1) gene of the cotton. In quantitative PCR assay, the quantification range was from 0.01 to 100% in 100 ng of the genome DNA template, and in the detection of 1.0, 3.0, and 5.0% levels of three insect resistant cotton lines, respectively, all of the relative standard deviations (RSDs) were less than 8.2% except for the GM cotton samples with 1.0% Mon531 or GK19, which meant that our real-time PCR assays involving the use of reference molecule were reliable and practical for GM insect resistant cottons quantification. All of these results indicated that our established conventional and TaqMan real-time PCR assays were applicable to detect the three insect resistant cottons qualitatively and quantitatively.

Animals↗

PubMed2003

Dimensions of novelty: a social representation approach to new foods.

Social representations of new foods were examined with a total of 44 subjects in nine focus groups. Each group was homogenous, defined by age, gender and educational background. Halfway through the interview, commercial packages of functional, genetically modified, organic, nutritionally modified and ethnic foods were presented as visual stimuli for discussion. Thematic and content analyses of the interview data showed that five dichotomies characterized the social representation: trust/distrust, safe/unsafe, natural/artificial, pleasure/necessity, and past/present. Many metaphors were used, with functional products being associated metaphorically with, for example, medicine and genetically modified products being associated with death and terrorism. Chronological references focused on the development of cuisine. The perceived unsafety of new foods was an important argument for women but not for men. The difference between age groups was in relating the discussion to either present time (young subjects) or past time (older subjects). Level of education affected the content of argumentation. In the context of new foods, social representations are formed to cope with the feeling of strangeness evoked by the novelties. They also have a role in cultural acceptance of new products by making them familiar. Overall, the results reflect the development of a new common sense in which popularized scientific notions are anchored in the process of urbanization.

Adolescent↗

PubMed2004

[Genetically modified plants and food safety. State of the art and discussion in the European Union].

Placing genetically modified (GM) plants and derived products on the European Union's (EU) market has been regulated by a Community Directive since 1990. This directive was complemented by a regulation specific for genetically modified and other novel foods in 1997. Specific labelling requirements have been applicable for GM foods since 1998. The law requires a pre-market safety assessment for which criteria have been elaborated and continuously adapted in accordance with the state of the art by national and international bodies and organisations. Consequently, only genetically modified products that have been demonstrated to be as safe as their conventional counterparts can be commercialized. However, the poor acceptance of genetically modified foods has led to a de facto moratorium since 1998. It is based on the lack of a qualified majority of EU member states necessary for authorization to place genetically modified plants and derived foods on the market. New Community Regulations are intended to end this moratorium by providing a harmonized and transparent safety assessment, a centralised authorization procedure, extended labelling provisions and a traceability system for genetically modified organisms (GMO) and derived food and feed.

Consumer Product Safety↗

PubMed2000

Implication of gene distribution in the bacterial chromosome for the bacterial cell factory.

As bacterial genome sequences accumulate, more and more pieces of data suggest that there is a significant correlation between the distribution of genes along the chromosome and the physical architecture of the cell, suggesting that the map of the cell is in the chromosome. Considering sequences and experimental data indicative of cell compartmentalisation, mRNA folding and turnover, as well as known structural features of protein and membrane complexes, we show that preliminary in silico analysis of whole genome sequences strongly substantiates this hypothesis. If there is a correlation between the genome sequence and the cell architecture, it must derive from some selection pressure in the organisms growing in the wild. As a consequence, the underlying constraints should be optimised in genetically modified organisms if one is to expect high product yields. Consequences in terms of gene expression for biotechnology are straightforward: knocking genes out and in genomes should not be randomly performed, but should follow the rules of chromosome organisation.

Bacteria↗

PubMed2003

Genes in food--why the furore?

Although unprecedented and perhaps unique in its irrationality, the recent furore over genetically modified (GM) food holds extremely important lessons for scientists. Some sections of the media undoubtedly bear a heavy responsibility for giving the expression 'GM' threatening connotations that are quite unwarranted. However, influential contributions to the hysteria have come from a surprising range of other sources, including some scientists. The research community has failed in its responsibility to society in three ways. Firstly, plant scientists did not appreciate that certain techniques (such as the use of antibiotic resistance genes as markers during plant transformation) would inevitably provoke public consternation. As a result, they took no steps to address such concerns. Secondly, researchers overlooked, minimized or in some cases simply dismissed the significance of public fears that they were 'interfering with Nature' or 'playing God'. Thirdly, plant breeders apparently saw no need to take pro-active measures with regard to the media and public in placing potential environmental and nutritional benefits of GM crops on the agenda in a positive fashion. Partly because of this failure, GM food is now firmly established in the public mind as wholly objectionable. One measure of how far we have travelled down that road is that it hardly matters any more whether objections are based on alleged environmental risks of cultivating GM crops or alleged toxicological hazards of eating them. 'Genetically modified organism', like 'radioactivity', has become an odious, generic shibboleth. Given that millions of people throughout the world are already benefiting from pharmaceuticals made by GM organisms, this is bizarre.

Communications Media↗

PubMed2006

Live bacterial vaccines--a review and identification of potential hazards.

The use of live bacteria to induce an immune response to itself or to a carried vaccine component is an attractive vaccine strategy. Advantages of live bacterial vaccines include their mimicry of a natural infection, intrinsic adjuvant properties and their possibility to be administered orally. Derivatives of pathogenic and non-pathogenic food related bacteria are currently being evaluated as live vaccines. However, pathogenic bacteria demands for attenuation to weaken its virulence. The use of bacteria as vaccine delivery vehicles implies construction of recombinant strains that contain the gene cassette encoding the antigen. With the increased knowledge of mucosal immunity and the availability of genetic tools for heterologous gene expression the concept of live vaccine vehicles gains renewed interest. However, administration of live bacterial vaccines poses some risks. In addition, vaccination using recombinant bacteria results in the release of live recombinant organisms into nature. This places these vaccines in the debate on application of genetically modified organisms. In this review we give an overview of live bacterial vaccines on the market and describe the development of new live vaccines with a focus on attenuated bacteria and food-related lactic acid bacteria. Furthermore, we outline the safety concerns and identify the hazards associated with live bacterial vaccines and try to give some suggestions of what to consider during their development.

Journal Article↗

PubMed1998

[Patents on life? No patent on life!].

Inventions related to living material are in principle patentable as well as inventions in the "classical" fields of technology as long as they are new, industrially applicable and involve an inventive step. A patent gives to its owner for a limited period of time the exclusive right to prevent others from using his patented new technical know-how. Starting point of patent protection in the field of genetic engineering is a genetic information or a genetically induced characteristic of an organism; there is no such thing as a "Patent on Life". As far as inventions relate to genetically modified organisms, patents give to their owners no additional property rights that might exclude the applicability e.g. of the laws on animal protection. Intellectual property like any other property is subject to the limits set up by law. It is neither scientifically correct nor does it help in finding a solution for the conflict within society to shift the--undoubtedly necessary--discussion about research and application in the field of genetic engineering to a discussion about patent law.

Animal Welfare↗

PubMed2005

What do individuals in different science groups within a life sciences organization think about genetic modification?

An assessment was undertaken of the attitudes of individuals within the science community towards a program to produce genetically modified cattle for altered milk composition, expectantly allowing for research into the treatment of multiple sclerosis in humans. The majority of respondents to an electronic survey expressed favorable attitudes to the program, thought it beneficial, respected individual freedom and was fair and just and disagreed that it was harmful. A passion for science and having a suitable lifestyle were the most important motivating factors for individuals. Finally, there were a wide range of responses to a number of cultural beliefs or myths. Science grouping significantly affected the responses. Compared with Systems and Land groups, Plant and Reproduction groups more strongly agreed with the project, thought it less harmful to interest groups, felt that genetic modification of animals was more morally acceptable, and more strongly agreed with the myth statements. These results indicate a diversity of beliefs and attitudes towards genetic modification amongst those within the science community, and highlight the importance of understanding ethics and myths in dealing with them. It is suggested that the diversity of beliefs could be better used to help shape public policy and understanding of biotechnology.

Agriculture↗

PubMed2001

Biotech prospects for the control of introduced mammals in Australia.

More than twenty exotic vertebrate species are now listed as pests in Australia. Collectively, these pests have a huge economic and environmental impact and pose a major threat to Australia's ecosystems and unique biodiversity. Management of such pests on a continental scale is a major challenge. Recent advances in biotechnology suggest alternatives to the lethal diseases normally sought for use as biological control agents. One proposal, being investigated in the Pest Animal Control Cooperative Research Centre, Canberra, is the use of biotechnology to develop a new generation of agents that act through controlling reproduction to prevent the build up of pest populations. The core concept is fertility control through immunocontraceptive vaccines delivered by viruses that specifically infect the target pest population. Proof of this exciting concept has been obtained for the mouse and, very recently, the rabbit, and a candidate vaccine vector identified for the fox, portending better control of a trio of Australia's most pervasive pests. Other advances in biotechnology suggest ways to negate the build up of both innate and acquired immune resistance in target pest populations that normally act to limit the efficacy and effective life of biocontrol agents in the field. Prospects for extending the use of virally vectored vaccines to the field management of wildlife diseases are also identified. Targets for such vaccines include a growing suite of emerging diseases, hosted by Australia's wildlife, which pose a threat to human and livestock health. Numerous technical challenges remain to be addressed before any of these new agents are ready for use in the field. However, the major risk to their development is now no longer viewed as being technical, but the failure to gain public acceptance for their use in the field. This already significant risk is exasperated by the present heightened level of public concern about all introductions of genetically modified organisms.

Animals↗

PubMed2006

Impact of Bt maize pollen (MON810) on lepidopteran larvae living on accompanying weeds.

Environmental risks of Bt maize, particularly pollen drift from Bt maize, were assessed for nontarget lepidopteran larvae in maize field margins. In our experimental approach, we carried out 3-year field trials on 6 ha total. Three treatments were used in a randomized block design with eight replications resulting in 24 plots: (i) near-isogenic control variety without insecticide (control), (ii) near-isogenic control variety with chemical insecticide (Baytroid) and (iii) Bt maize expressing the recombinant toxin. We established a weed strip (20 x 1 m) in every plot consisting of a Chenopodium album (goosefoot)/Sinapis alba (mustard) mixture. In these strips we measured diversity and abundance of lepidopteran larvae during maize bloom and pollen shed. C. album hosted five species but all in very low densities; therefore data were not suitable for statistical analysis. S. alba hosted nine species in total. Most abundant were Plutella xylostella and Pieris rapae. For these species no differences were detected between the Bt treatment and the control, but the chemical insecticide treatment reduced larval abundance significantly. Conclusions regarding experimental methodology and results are discussed in regard to environmental risk assessment and monitoring of genetically modified organisms.

Animals↗

PubMed1989

[Gene technology and biological safety].

A "European Biosafety Workshop on the Uncontained Use of Modified Organisms in the Environment" was held in Berlin on 27-30 November 1988. The author's concluding remarks are given here in a free German translation. The article briefly summarizes the overall conditions for a general evaluation of potential benefits and risks that may be associated with the use of genetically modified organisms.

Accident Prevention↗

PubMed2002

Hazard identification and risk assessment procedure for genetically modified plants in the field--GMHAZID.

The safe application of genetically modified organisms (GMOs) requires a risk assessment prior to their proposed use. Based on methods from the chemical industry, we developed a hazard identification procedure for the risk assessment of field tests with genetically modified plants. This risk assessment method, GMHAZID, is carried out in the form of guided brainstorm sessions. GMHAZID was tested with a case study for which a risk assessment had previously been made, and the results of the assessments were compared. The results showed that some new hazards potentially leading to uncontrolled spreading, in addition to those from the previous assessment, were identified using GMHAZID. GMHAZID also recognised some hazards leading to failures in the field experiments. We suggest that GMHAZID provides systematics, reliability, and transparency to the risk assessment procedure.

Environmental Monitoring↗

PubMed2004

[Identification for genetically modified maize T14/T25 with real time fluorescent PCR method].

To identify genetically modified (GM) maize T14/T25 lines, a real-time fluorescent PCR (RTF PCR) assay was performed in this study. Primers and Taqman probes specific for inserted genes in the T14/T25 were used to conduct the real-time fluorescent (RTF) PCR and PCR assays. The RTF PCR method was established to detect and identify GM maize lines. The results show that the TaqMan probe could identify T14/T25 maize used, while other GM and NO-GM maize didn't be detected. The RTF PCR could be a new method for detecting other genetically modified organism.

DNA Primers↗

PubMed2001

Production of polyhydroxyalkanoates from intact triacylglycerols by genetically engineered Pseudomonas.

Pseudomonas putida and P oleovorans have been extensively studied for their production of medium-chain-length (mcl)-polyhydroxyalkanoates (PHA). These bacteria are incapable of metabolizing triacylglycerols (TAGs). We have constructed recombinant P. putida and P. oleovorans that can utilize TAGs as substrates for growth and mcl-PHA synthesis. A recombinant plasmid, pCN51lip-1, carrying Pseudomonas lipase genes was used to electrotransform these organisms. The transformants expressed TAG-hydrolyzing activity as shown by a rhodamine B fluorescence plate assay. The genetically modified organisms grew in TAG-containing medium to a cell dry weight of 2-4 g/l. The recombinant P. putida produced mcl-PHA at a crude yield of 0.9-1.6 g/l with lard or coconut oil (Co) as substrate. While P. oleovorans transformant did not produce mcl-PHA, a mixed-culture fermentation approach with the wild-type and recombinant strains afforded polymer production from Co at a crude yield of 0.5 g/l. Compositional analysis by gas chromatography/mass spectrometry showed that beta-hydroxyoctanoate (31-45 mol %) and beta-hydroxydecanoate (28-35 mol %) were the dominant repeat units of the TAG-based PHA. The number-average and weight-average molecular masses of the PHAs as determined by gel permeation chromatography were 82-170 x 10(3) g/mol and 464-693 x 10(3) g/mol, respectively. The recombinant approach can greatly increase the number of organisms that can be used to produce PHA from fat and oil substrates.

Culture Media↗

PubMed2002

Experience with environmental issues in GM crop production and the likely future scenarios.

In the Cartagena Protocol on Biosafety, standards for risk assessment of genetically modified organisms (GMOs) have been set. The criteria and information basis for the risk assessment of GMOs have been modified by the EU Directive 2001/18/EC. Various approaches to further improve the criteria for environmental risk assessment of GMOs are described in this study. Reports on the ecological impacts of the cultivation of certain non-transgenic crop plants with novel or improved traits as analogy models to transgenic plants showed that the effects of agricultural practice can be at least equally important as the effects of gene transfer and invasiveness, although the latter currently play a major role in risk assessment of transgenic crops. Based on these results the applicability of the methodology of 'Life Cycle Analysis (LCA)' for genetically modified plants in comparison with conventionally bred and organically grown crop plants was evaluated. The methodology was regarded as applicable with some necessary future improvements. In current projects, the assessment of toxicology and allergenicity of GM crops are analysed, and suggestions for standardization are developed. Based on results and recommendations from these efforts there are still the challenges of how to operationalize the precautionary principle and how to take into account ecologically sensitive ecosystems, including centres of origin and centres of genetic diversity.

Agriculture↗

PubMed2025

Innovations in Transgene Integration Analysis: A Comprehensive Review of Enrichment and Sequencing Strategies in Biotechnology.

Understanding the integration of transgene DNA (T-DNA) in transgenic crops, animals, and clinical applications is paramount for ensuring the stability and expression of inserted genes, which directly influence desired traits and therapeutic outcomes. Analyzing T-DNA integration patterns is essential for identifying potential unintended effects and evaluating the safety and environmental implications of genetically modified organisms (GMOs). This knowledge is crucial for regulatory compliance and fostering public trust in biotechnology by demonstrating transparency in genetic modifications. This review highlights recent advancements in T-DNA integration analysis, specifically focusing on targeted DNA enrichment and sequencing strategies. We examine key technologies, such as polymerase chain reaction (PCR)-based methods, hybridization capture, RNA/DNA-guided endonuclease-mediated enrichment, and high-throughput resequencing, emphasizing their contributions to enhancing precision and efficiency in transgene integration analysis. We discuss the principles, applications, and recent developments in these techniques, underscoring their critical role in advancing biotechnological products. Additionally, we address the existing challenges and future directions in the field, offering a comprehensive overview of how innovative DNA-targeted enrichment and sequencing strategies are reshaping biotechnology and genomics.

Transgenes↗