Browsing by Author "Pando Tacuri, Heydi Isabel"

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Producción de las proteínas de choque térmico bovinas HSPA1A y HSPA2 en Escherichia coli
(Universidad de Cuenca, 2024-02-06) Largo Maldonado, Estefany Mishel; Pando Tacuri, Heydi Isabel; Vallecillo Maza, Antonio Javier
Chaperone proteins are responsible for maintaining the quality and functionality of other proteins. Some chaperones are involved in various functions within reproductive physiology, such as maintaining the quality and functionality of gametes, among which are chaperones HSP70 family, which are expressed mainly in response to stressful situations such as heat shock, oxidative and osmotic stress, among others. Furthermore, members of this family have also been identified in the plasma membrane of the sperm and decreases in their expression compromise their motility and quality. In view of this background, this work focused on the production of recombinant proteins HSPA1A and HSPA2 from bovines (Bos taurus) for their evaluation as an additive in cryopreservation solutions for embryos and gametes. As a result, it was only possible to obtain the HSPA1A protein in a bacterial expression model; with the Escherichia coli (E. coli) Rosetta 2 (DE3) strain, approximately 112 mg of pure protein per liter was obtained, in cultures incubated at a temperature of 37 °C. On the contrary, in production cultures incubated at a temperature of 30 °C, a yield of ± 6 mg of pure protein was achieved. On the other hand, it was not possible to achieve the production of the HSPA2 protein, as a result of being able to amplify the coding sequence, so another amplification strategy must be designed and thus be able to obtain the genetic construction necessary for the generation of the recombinant strain of E. coli capable of producing this protein.