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Genetic Modification

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Introduction

In a comment to the press that the EU Commission gave out together with the report this week they claimed that "there is nothing secret about the study referred to in the Greenpeace press release. The version published on their [the Greenpeace] web-site is a draft ..." However, this is simply not true. The letters that accompanied the study when it was delivered to the European Commission in January, and that were obtained by Greenpeace, clearly state that the study was presented in its "final version" at that time. The study states that farmers who don't want to cultivate GMOs would face high additional, in some cases unsustainable, costs of production if genetically engineered (GE) crops were commercially grown on a large scale in Europe. The study predicts that the situation would become particularly critical for organic farming of oilseed rape as well as for intensive production of conventional maize. Seed and crop purity from GE pollution, at a detection level of 0.1 percent, would be virtually impossible in most cases. This effectively means that all products and seeds of oilseed rape and maize would be contaminated with GE crops to a certain extent. Organic farming exempt of GMOs, as we know it today and as it is defined in the EU Regulations, will be doomed. These findings confirm the need for "zero tolerance" for seed contamination, the standard demanded by Greenpeace and other organisations Over the past few years, many novel human genes have been discovered by means of systematic DNA sequencing projects. For some of these genes it is possible to infer a probable function by comparative sequence analysis, however, for many others this is not possible and their functions remain unknown. Some of these genes could be responsible for genetic neuromuscular diseases and cardiomyopathies that are still uncharacterised at the molecular level; the aim of the Muscle Molecular Biology (MMB) ...read more.

Middle

To identify origin localisation in mega-base regions of the human genome, we have now developed a procedure for long-range origin mapping. The method (Subtractive Hybridisation for Origin Capture, SHOC assay) is based on the subtractive hybridisation of a nascent DNA library to contigs from the genomic region of interest. The selected DNA fragments are representatives of origins belonging to the analysed region and can be used as hybridisation probes against the cosmids which drove the selection. In collaboration with H. Masai, University of Tokyo, Japan, the SHOC assay was used for the identification of origins in a ~100 kb region of the long arm of human chromosome 5, in a region containing the IL3 and GM-CSF genes. The results indicated the presence of two origins in this area, spaced ~30 kb apart and located ~20 kb upstream of the IL3 gene and ~5 kb downstream of the GM-CSF gene. Analysis of these origins, as well as of other, already characterised ones, including the lamin B2 origin, suggests the existence of a conserved consensus sequence in the core origin region. This core sequence is the cis-acting substrate for protein binding in vivo, as revealed by genomic footprinting experiments. Interaction of a novel origin in the 5q area, as well as of the already characterised lamin B2 origin, with DNA replication proteins was analysed using quantitative chromatin immunoprecipitation. The results obtained with antibodies against hOrc2, hCDC6, and hMcm5 are suggestive of cell cycle-regulated recruitment of these factors to the core origin sequences. Gene therapy Gene transfer using Adeno-Associated Virus (AAV) vectors Due to the absence of pathogenicity, the ease of production of high titer preparations, and the apparently, low genetic complexity, vectors based on AAV, a replication-defective parvovirus, represent very interesting tools for gene transfer. In particular, rAAV vector preparations are highly successful in transducing and expressing reporter genes in neuronal and muscular cells in vivo. ...read more.

Conclusion

MAGI regulation of the PTEN phosphatase To date, there is no evidence that Dlg or the MAGI family of proteins are mutated during the development of human cancers and hence cannot be classed as true tumour suppressor proteins. However, both of them have been shown to interact with, and regulate, the activity of known tumour suppressor proteins; Dlg with APC and MAGI with the PTEN phosphatase. Figure 3 shows a schematic of the Protein Kinase B (PKB) pathway which is regulated by the PTEN phosphatase and the MAGI family of proteins. As can be seen, blocking MAGI activity by E6 will result in reduced PTEN activity and increased signalling from PKB. Studies are now in progress to investigate these possibilities in more detail and, in particular, whether another HPV oncoprotein, E5, may also impact on this pathway, since we have previously shown that E5 can activate signalling from a variety of growth factor receptors. Regulation of Viral DNA replication We previously reported that the viral transcriptional regulator, E2, can also interact with p53. This activity appeared to correlate with the ability of p53 to inhibit viral DNA replication. A major outstanding question however, is why this interaction takes place. Recent reports have indicated that the viral capsid protein, L2, colocalises with p53 and also interacts with E2. These results suggest a complex interplay between these proteins during viral DNA replication and subsequent DNA packaging into the viral capsid. To investigate these phenomena in more detail we have initiated a series of studies to map the E2-L2 interaction domains. At the same time, we will also use a panel of L2 mutants to determine which sequences determine L2 subcellular location, and we also propose to investigate the role of p53 in these processes. Figure 3. Regulation of Protein Kinase B (PKB) by the PTEN phosphatase and MAGI. Activation of PI'3 Kinase (PI3K) following growth factor activation phosphorylates phosphatidyl inositols (PtdIns) which in turn activates PKB. This is downregulated by the PTEN/MAGI complex, which dephosphorylates the PtdIns. The putative inhibitory effect of E6 on this regulation is also shown. ...read more.

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