Browsing by Author "Arce Medina, Aníbal Andrés"

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    CFE Expression and Lyophilization of β-Galactosidase (LacZ) v1
    (2022) Navarro Martínez, Felipe; Arce Medina, Aníbal Andrés; Federici, Fernan
    This protocol includes the expression and lyophilization of the β-Galactosidase enzyme (from the LacZ gene) in a cell-free system. We used it to prepare and conduct experiments for a biochemistry lab course.
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    Omics Approaches for Understanding Grapevine Berry Development: Regulatory Networks Associated with Endogenous Processes and Environmental Responses
    (2017) Serrano, A.; Espinoza, C.; Armijo, G.; Inostroza, C.; Poblete, E.; Meyer Regueiro, Carlos José; Arce Medina, Aníbal Andrés; Parada, F.; Santibanez, C.; Arce Johnson, Jorge Patricio
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    Open platform for the implementation of RNA sensing reactions in cell-free systems
    (2021) Arce Medina, Aníbal Andrés; Federici, Fernán; Pontificia Universidad Católica de Chile. Facultad de Ciencias Biológicas
    La biología sintética busca el desarrollo de funciones programables y predecibles en sistemas biológicos. Uno de los recientes avances en esta área son los toehold swtiches: reguladores de la expresión genética diseñados denovo y que permiten la traducción de un gen de forma gatillada por la interacción con un RNA trigger" de secuencia especifica. Por otro lado, los sistemas de expresión genética libre de células (cell-free) han sido utilizados durante décadas en el campo de la biología molecular y fueron claves en el descubrimiento del código genético. Sin embargo, solo muy recientemente han empezado a utilizarse como una herramienta para la bioingeniería. La expresión de toehold swtiches en sistemas de libre de células generó una plataforma de diagnóstico muy promisoria ya que los sensores de RNA tipo toehold pueden ser liofilizados para su transporte a temperatura ambiente con el objetivo de ser utilizadas en terreno y en zonas remotas. No obstante, a la fecha, estos sensores solamente han sido implementados en sistemas de expresión genética reconstituidos, del tipo PURE (por sus siglas en inglés: Protein expression Using Reconstituted Elements). Los sistemas PURE requieren transporte en fr´ıo (-80 C) desde los proveedores en el hemisferio norte, y son prohibitivamente caros para los usuarios en América Latina.
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    Regulation of polar auxin transport in grapevine fruitlets (Vitis vinifera L.) and the proposed role of auxin homeostasis during fruit abscission
    (2016) Kühn Weber, Nathalie Alexandra; Serrano Canales, Alejandra Rita de Lourdes; Abello, Carlos; Arce Medina, Aníbal Andrés; Espinoza Cancino, Carmen Gloria; Gouthu, Satyanarayana; Deluc, Laurent; Arce Johnson, Jorge Patricio
    Background Indole-3-acetic acid (IAA), the most abundant auxin, is a growth promoter hormone involved in several developmental processes. Auxin homeostasis is very important to its function and this is achieved through the regulation of IAA biosynthesis, conjugation, degradation and transport. In grapevine, IAA plays an essential role during initial stages of berry development, since it delays fruitlet abscission by reducing the ethylene sensitivity in the abscission zone. For this reason, Continuous polar IAA transport to the pedicel is required. This kind of transport is controlled by IAA, which regulates its own movement by modifying the expression and localization of PIN-FORMED (PIN) auxin efflux facilitators that localize asymmetrically within the cell. On the other hand, the hormone gibberellin (GA) also activates the polar auxin transport by increasing PIN stability. In Vitis vinifera, fruitlet abscission occurs during the first two to three weeks after flowering. During this time, IAA and GA are present, however the role of these hormones in the control of polar auxin transport is unknown. Results In this work, the use of radiolabeled IAA showed that auxin is basipetally transported during grapevine fruitlet abscission. This observation was further supported by immunolocalization of putative VvPIN proteins that display a basipetal distribution in pericarp cells. Polar auxin transport and transcripts of four putative VvPIN genes decreased in conjunction with increased abscission, and the inhibition of polar auxin transport resulted in fruit drop. GA3 and IAA treatments reduced polar auxin transport, but only GA3 treatment decreased VvPIN transcript abundance. When GA biosynthesis was blocked, IAA was capable to increase polar auxin transport, suggesting that its effect depends on GA content. Finally, we observed significant changes in the content of several IAA-related compounds during the abscission period. Conclusions These results provide evidence that auxin homeostasis plays a central role during grapevine initial fruit development and that GA and IAA controls auxin homeostasis by reducing polar auxin transport.
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    Step 2: Preparing amino acid, polyphosphates, and maltodextrin-based energy solutions for cell-free reactions V.3
    (2018) Arce Medina, Aníbal Andrés; Puig Contreras, Juan Manuel; Federici, Fernán