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La ingeniería genética y la terapia génica. Su relación con la ética y el derechoOsorio Osorio, Francisco Antonio, Pinto Arqueros, Yurit Patricio January 2000 (has links)
Memoria (licenciado en ciencias jurídicas y sociales) / No autorizada por el autor para ser publicada a texto completo / La presente memoria de prueba tiene como centro de atención a la ingeniería genética y en especial a la terapia génica como expresiones de la biotecnología y biomedicina aplicada a humanos, nos proponemos reflexionar desde una perspectiva filosófica, Bioética y sobretodo jurídica sobre estos problemas de tanta actualidad y futuro, especialmente si tomamos en cuenta que en nuestro país estos temas no se encuentran normativamente integrados en nuestra legislación; por lo tanto, pretende ser un aporte en la dirección de: informar, reflexionar y en definitiva poder regular jurídicamente los diversos aspectos que involucra. Hemos preparado esta memoria de prueba teniendo presente en especial la perspectiva que nos otorga la disciplina bioética, observando también la legislación extranjera en esta materia y contando en todo momento con la colaboración inestimable de nuestro profesor guía don José Miguel Vera Lara.
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Manejo de la producción de salinosporamide A en salinispora trópica CNB-440 empleando ingeniería metabólica y genéticaSaucedo Hernández, Vianey Diana January 2019 (has links)
Tesis para optar al grado de Doctora en Ciencias de la Ingeniería Mención Ingeniería Química y Biotecnología / Salinosporamide A is a cytotoxic that has been proven to combat various types of cancer and malaria. It is currently in phases II and III of approval as an anticarcinogen. The advantages that poses over other cytotoxics are greater activity at low concentrations and highly specific. It acts on the proteasome-ubiquitin system, responsible of apoptosis in cells.
This secondary metabolite is naturally ocurred in the actinomycetes bacterium strictly marine, Salinispora tropica that needs a specific ionic force in the medium to grow. Due to his nature it is a promisory source of secondary metabolites for pharmaceutical use hereby is constantly studied. The CNB440 strain is the representative strain of the species and posses a Genome-Scale Metabolic Model (GSM). The goal of this work was to implement diverse metabolic and genetic strategies that allow improve the production of Salinosporamide A.
Chapter 4 of this thesis details the proves to establish the protocols for growth and determination of Salinosporamide A, Define the sensitivity of bacteria to kanamycin (100 ug/ml), thiostreptone (12 µg/ml) and apramycin (12 µg/ml). The growth curves in several minimum mediums, and stablish the methodology for the determination of Salinosporamide A.
Chapter 5 describes the genetic strategy used to modify the bacterium and generate a higher concentration of Salinosporamide A. The strategy followed was by recombination homologous with the temperature sensitive vector (pGM1190) and transferred to S. tropica by conjugation with the strain E. coli ET12567/pUZ8002, to delete specific sites on the chromosome of S. tropica. These molecular tools have been successfully used in the transformation of various Streptomyces, but had not been tested in Salinispora. The sites suggested to be deletedto increase the production of the secondary metabolite were 3 clusters of genes sporolides, lymphostine and salinilactam. But due to various complications in the development of the present work only the deletion of the sporolide gene cluster was evaluated and this resulted in an increase of 20% in metabolite production.
Chapter 6 details the use of genome-scale metabolic model iCC908 for increase the production of Salinosporamide A. The first stage consisted in establishing the working environment of the model, to increase the accuracy of the model, integrated growth data, metabolites in medium production and determination of Salinosporamide A, with this was also able define in silico the supplementation of medium production, to obtain more Salinosporamide A. . The second stage consisted of applying different algorithms OptKnock, OptGene, OptOrf, GDLS, FSEOF, which browse reactions or genes within the genome-scale model that could be, deleted, blocked or overexpressed to increase the production of the secondary metabolite. We found several candidates that were evaluated in silico and we proposed to evaluate the deletion of two genes. As the last stage, were evaluated the metabolic pathways that increase production by gene overexpression. The evaluation of this metabolic pathways consist in add diverse substrates that increase the flow in the pathway of the gene to be overexpresed, tyrosine at a concentration of 5mM increase the production of the secondary metabolite Salinosporamide A by 180%, enhancing the presence of phenylalanine in the medium.
With these results it was possible to obtain a medium production that increased in 2.8 times Salinosporamide A by fermentation based on the use of the genome-scale metabolic model. And also was possible transform the strain S. tropica with genetic tools previously proved in Streptomyces.
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