70 resultados para transformation gfp


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It is critical that viruses are able to avoid the antiviral activities of interferon (IFN). We have shown previously that the human papillomavirus (HPV) is able to avoid IFN-alpha via interaction of the HPV-16 E7 protein with IFN regulatory factor-9 (IRF-9). Here, we investigated the details of the interaction using HPV-16 E7 peptide mapping to show that IRF-9 binds HPV-16 E7 in a domain encompassing amino acids 25-36. A closer examination of this region indicates this is a novel proline, glutamate, serine, and threonine-rich (PEST) domain, with a PEST score of 8.74. We have also mapped the region of interaction within IRF-9 and found that amino acids 354-393 play an important role in binding to HPV-16 E7. This region of IRF-9 encompasses the IRF association domain (IAD), a region important for protein-protein interaction central to IRF function. Finally, we used alanine-scanning mutagenesis to determine if E7-IRF-9 interaction was important for E7-mediated cellular transformation and found that the HPV-16 E7 mutants Y25A, E26A, S31A, S32A, and E35A, but not L28A and N29A, caused loss of transformation ability. Preliminary data suggest loss of IRF-9 interaction with E7 mutants correlated with transformation. Our work suggests E7- IRF- 9 interaction is important for the transforming ability of HPV-16 E7 and that HPV-16 E7 may interact with other IRF proteins that have IAD domains.

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A fast, reproducible, and efficient transformation procedure employing Agrobacterium rhizogenes was developed for Phaseolus vulgaris L. wild accessions, landraces, and cultivars and for three other species belonging to the genus Phaseolus: R coccineus, P lunatus, and P acutifolius. Induced hairy roots are robust and grow quickly. The transformation frequency is between 75 and 90% based on the 35-S promoter-driven green fluorescent protein and beta-glucuronidase expression reporter constructs. When inoculated with Rhizobium tropici, transgenic roots induce normal determinate nodules that fix nitrogen as efficiently as inoculated standard roots. The A. rhizogenes-induced hairy root transformation in the genus Phaseolus sets the foundation for functional genomics programs focused on root physiology, root metabolism, and root-microbe interactions.

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This study examined employee readiness for fine-tuning changes and for corporate transformation changes. It was proposed that employees would report different degrees of readiness for these two types of change and that different variables would be associated with readiness for the two types of change. Results of regression analyses indicated that trust in peers and logistics and system support displayed strong positive relationships with readiness for fine-tuning changes, while trust in senior leaders and self-efficacy displayed strong positive relationships with readiness for corporate transformation changes. The implications of this study focus on the appropriateness of traditional change management strategies in light of findings that multiple change readiness attitudes exist within an organization.

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Civil society is recognised as comprising complex and multifaceted entities, resilient to and yet responsive to both the state apparatus and global market processes. Civil society in the Philippines, long regarded as one of the most vibrant, diverse and innovative in Asia, has emerged as a significant actor in the field of conflict resolution and peace-building. In thinking about the work of peace, this paper engages with the effectiveness of civil society in mobilising societal awareness for a ‘just and lasting peace’ in the southern Philippines. Shaped by development paradigms that privilege concepts such as social capital, the paper aims to interrogate how such concepts situated within the development–security nexus proposed by the Philippine government and funding agencies have influenced conflict-transformation initiatives in Mindanao, Philippines.

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Model transformations are an integral part of model-driven development. Incremental updates are a key execution scenario for transformations in model-based systems, and are especially important for the evolution of such systems. This paper presents a strategy for the incremental maintenance of declarative, rule-based transformation executions. The strategy involves recording dependencies of the transformation execution on information from source models and from the transformation definition. Changes to the source models or the transformation itself can then be directly mapped to their effects on transformation execution, allowing changes to target models to be computed efficiently. This particular approach has many benefits. It supports changes to both source models and transformation definitions, it can be applied to incomplete transformation executions, and a priori knowledge of volatility can be used to further increase the efficiency of change propagation.