Examinando por Autor "Abaunza Villamizar, Sebastián Mauricio"

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    Acceso abierto
    Caracterización de nuevas proteínas Cry11 obtenidas por modelos heurísticos computacionales
    (Bucaramanga : Universidad de Santander, 2018, 2018-11-23) Abaunza Villamizar, Sebastián Mauricio; Suárez Barrera, Miguel Orlando
    Bacillus thuringiensis (Bt) is a Gram positive bacterium with parasporal inclusion bodies, also called δ-endotoxins, within these is the Cry protein that is known for its toxic activity against multiple orders of insects and nematodes, the vector-borne diseases A. aegypti, it transmites several important diseases in public health such as Dengue, Zika Chikungunya, among others. Cry11 has been recognized for its high specificity against A. aegypti. However, the emergence of resistance by insects that are the object of study, have resulted in the application of various strategies, for the improvement of proteins including site-directed mutagenesis and DNA shuffling. The use of biocomputational tools has expanded the spectrum in the development of molecules with greater potential, however, these strategies have not been described in the development of enhanced mutants of Cry. This work is based on the use of software HIDDEN 1.0 where libraries of in silico variants were obtained from Cry11Aa. Heu2, Heu3 and Heu4 variants were selected, and validated in vitro, they were synthesized, cloned into PSV2 and were expressed in E.coli DE3BL21 and B. thuringiensis BMB171, their electrophoretic profiles were analyzed in SDS-Page and their toxicity activity was evaluated by thick tests with A. aegypti. Parallel to this a structural analysis with matrix of MatGat, Bioedit and structural comparisons were contrasted with the literature. It was found that the variants presented an upper identity more than 97% with Cry11Aa, the changes mostly belonged to domain II, and their lethality percentages were less than 5%, which suggests that the amino acid changes in the important regions of the three domains are involved in the toxicity activity demonstrated by the proteins.