Construcción molecular del vector pTNR-KA-DsRed para el marcaje fluorescente de la cepa C-924
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Fecha
2019
Autores
Díaz González, Omar
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Universidad Central ``Marta Abreu`` de Las Villas. Facultad de Ingeniería Química y Farmacia. Departamento de Licenciatura Química
Resumen
Los análisis de la interacción entre las plantas hospedantes y las bacterias beneficiosas en su entorno natural son esenciales para la elucidación de su funcionamiento y la aplicación exitosa de agentes comerciales de control biológico. El uso de proteínas fluorescentes como marcadores permite una caracterización completa de la interacción planta-microbioma "in situ". El presente trabajo se desarrolla en el Centro de Ingeniería Genética y Biotecnología de Camagüey (CIGB), que actualmente en el área de investigaciones trabaja en el marcaje molecular de la cepa bacteriana C-924, principio activo del bionematicida Hebernem-S®. Entre las plagas que afectan el sector agrícola y ganadero, los nematodos son responsables de pérdidas económicos considerables. Actualmente el uso de bioproductos constituye una alternativa para el control de esta plaga. El objetivo de este trabajo es obtener un vector de expresión capaz de transformar la cepa C-924 con el gen fluorescente DsRed mediante técnicas moleculares. Se obtuvo la construcción pNit-DsRed-thcA potencialmente funcional a partir del análisis “in sillico” de la secuencia nucleotídica y aminoacídica. Además, se logró obtener el vector pTNR-KA-DsRed que constituye un elemento crucial para el marcaje fluorescente de la cepa C-924.
Interaction`s analysis between host plants and beneficial bacteria are essential for the elucidation of their functioning and the successful commercial biological control agents`s application. Fluorescent proteins`s use as markers allows a complete characterization of the plant-microbiome interaction "in situ". This work is carried out at the Center for Genetic Engineering and Biotechnology of Camagüey (CIGB), which currently works in the molecular labeling of strain C-924, the active ingredient of the bionematicide Hebernem-S®. Among the pests that affect the agricultural and livestock sector, nematodes are responsible for considerable economic losses. Currently the use of bioproducts is an alternative for the control of this pest. The aim of this work is to obtain an expression vector capable of transforming the strain C-924 with the fluorescent gene DsRed by means of molecular techniques. The potentially functional pNit-DsRed-thcA construct was obtained from the "in sill" analysis of the nucleotide and amino acid sequence. In addition, it was possible to obtain the pTNR-KA-DsRed vector, which is a crucial element for the fluorescent labeling of strain C-924.
Interaction`s analysis between host plants and beneficial bacteria are essential for the elucidation of their functioning and the successful commercial biological control agents`s application. Fluorescent proteins`s use as markers allows a complete characterization of the plant-microbiome interaction "in situ". This work is carried out at the Center for Genetic Engineering and Biotechnology of Camagüey (CIGB), which currently works in the molecular labeling of strain C-924, the active ingredient of the bionematicide Hebernem-S®. Among the pests that affect the agricultural and livestock sector, nematodes are responsible for considerable economic losses. Currently the use of bioproducts is an alternative for the control of this pest. The aim of this work is to obtain an expression vector capable of transforming the strain C-924 with the fluorescent gene DsRed by means of molecular techniques. The potentially functional pNit-DsRed-thcA construct was obtained from the "in sill" analysis of the nucleotide and amino acid sequence. In addition, it was possible to obtain the pTNR-KA-DsRed vector, which is a crucial element for the fluorescent labeling of strain C-924.
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Centro de Ingeniería Genética y Biotecnología(CIGB), Camagüey, Ingeniería Genética, Biología Molecular, Reacción en Cadena de la Polimerasa (PCR), Cepa Bacteriana C-924, Vector pTNR-KA-DsRed