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Espinosa M. Escuela de Administraci. Greenwich, CT: Inforrnation Age Publishing Inc. Contenido. Introducci. Inteligencia emocional, liderazgo transformacional y g. El rol de la mujer en el liderazgo / Andrea Patricia Cuestas D. Liderazgo transformacional, clima organizacional, satisfacci. La mujer en la alta direcci. Influencia de la personalidad en el discurso y liderazgo de George W. El liderazgo desde la perspectiva del poder y la influencia. El trabajo directivo para l. Emotional intelligence and its relationship with leadership practices. International Journal of Business and management, 5(2), 6. Citation. Arteaga, A., & R. Gerencia Cognicional: la integraci. Revista de Estudios Interdisciplinarios en Ciencias Sociales, 1. Citation. Avolio, F. Leadership: Current Theories, Research, and Future Directions. Citation. Annu. Psychol, 6. Citation. Barling, J., Slater, F., & Kelloway, E. Transformational leadership and emotional. Leadership and organization development journal, 2. 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Aggressive behavior, 4. Citation. Wille, B. Fifty Shades of Personality: Integrating Five- Factor Model Bright and Dark Sides of Personality at Work. Industrial and organizational psychollogy. Citation. Zakaria, F. Washington Post. dc. Citation. Baker, S. The Theorical Foundation of Contemporary Construct. Journal of Leadership & Organizational Studies. Citation. Baker, S. Die Sims 2: Cheats und Tipps (PC, NDS, PS2, PSP, XBox, GBA, GC, Mac)Komplettl? Mit diesen Tipps legt ihr einen guten Start hin! Seiten Jede Menge Tipps f.
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A morpholino, also known as a morpholino oligomer and as a phosphorodiamidate morpholino oligomer (PMO), is type of oligomer molecule (colloquially, an oligo) used in molecular biology to modify gene expression. Hollstein et al., 1. Levine et al., 1. Greenblatt et al., 1. In many human tumours, the mutant p. The p. 53 gene is highly conserved in evolution, and has been studied in a wide range of species from human to flies and worms (Soussi and May, 1. Jin et al., 2. 00. Sutcliffe and Brehm, 2. The zebrafish is an excellent vertebrate model for embryological and genetic analysis, and is now the most widely used teleost species in biomedical research. In combination with its use in carcinogenesis assays (Stanton, 1. Stern and Zon, 2. Studies have shown that exposure of zebrafish to carcinogens induces a significant number and variety of neoplasms (Spitsbergen et al., 2. From a histopathology point of view, the fish neoplasms are remarkably similar to human cancer (Spitsbergen and Kent, 2. However, the use of the zebrafish as a model organism in p. Previous work has mainly been dependent upon in situ hybridization of m. RNA to study gene expression and apoptosis detection by TUNEL labeling (Langheinrich et al., 2. The availability of antibodies against zebrafish p. In this present study, we describe the production and characterization of monoclonal antibodies directed against zebrafish p. We demonstrate that the monoclonal antibodies are useful for immunoblotting and immunohistochemistry in zebrafish embryos. We show that intense nuclear p. Top of page. Results. Purification of zebrafish p. To generate the recombinant antigen for immunization, tp. DNA of the zebrafish (Danio rerio) was generated by PCR and expressed with a histidine tag in Escherichia coli. The use of the Overnight Express Autoinduction system (Novagen) permitted high levels of His- p. Figure 1, lane 2). Cell fractionation studies, however, show that the His- p. Figure 1a, lane 4) from which it could be purified (Figure 1a, lane 5). MALDI mass spectroscopy analysis of this band showed a high return score for zebrafish p. Huntington disease iPSCs show early molecular changes in intracellular signaling, the expression of oxidative stress proteins and the p53 pathway.The molecular weight size of 5. Da contrasts significantly with the calculated mass of 4. Da based on the amino acid sequence of zebrafish p. Oren, 1. 98. 5). Expression of recombinant zebrafish p. Escherichia coli followed by generation and characterization of p. Aliquots of purification extracts separated by electrophoresis on 4–1. SDS gel show whole cell lysates containing p. ET1. 9b (lane 1) and p. ET1. 9b- p. 53 (lane 2), soluble protein extract following Ni. Ni. 2+- charged chromatography purification (lane 5). Arrow indicates the position of the p. The monoclonal antibody ZFp. H1. 29. 9 cells (left panel), whereas strong cell staining was seen on sections of p. H1. 29. 9 cells (right panel). ZFp. 53- 5. 1 (lane 2) and ZFp. ZF- R1. 8 (lanes 1–4). Similarly, ZF- R1. ZFp. 53- 5. 1 (lanes 5 and 6) and probed by ZFp. Control samples (lanes 1, 3, 5 and 7) immunoprecipitated with an antibody to haemaglutinin (HA) are negative when immunoblotted with ZF- R1. ZFp. 53- 5. 1 (lane 5) or ZFp. Mr values of the molecular weight markers are indicated on the left (k. Da). Full figure and legend (8. K)Production and screening of the anti- p. A total of 9. 00 mouse hybridoma colonies were obtained after the fusion of splenocytes from immunized mice and myeloma cells and these were subjected to rigorous screening designed to identify antibodies suitable for immunohistochemistry on formalin- fixed paraffin- embedded tissues. After initial screening, 4. H1. 29. 9 cells, stained as sections of cell pellets fixed in formalin and embedded in agarose. Supernatant from 1. Figure 1b), and these were single- cell cloned and further analysed. H1. 29. 9 cell lysates were immunoprecipitated with monoclonal antibodies ZFp. ZFp. 53- 9. 1 and a negative control antibody to a haemagglutinin (HA) epitope tag. Immunoblots of these immunoprecipitates were then probed with a rabbit anti- zebrafish p. ZF- R1. 8, also generated in this study. Similarly, ZF- R1. ZFp. 53- 5. 1, ZFp. HA epitope antibody. The results show that the monoclonal antibodies and the ZF- R1. Figure 1c), thus demonstrating the sensitivity and specificity of zebrafish p. Using a set of overlapping zebrafish p. Figure 2). Of the 1. Ig. G isotype while the remaining four were Ig. M isotype (Table 1, column 2). The antibodies were also tested for cross- reactivity and specificity on four human breast cancer cell lines. The BT4. 74, BT5. T4. 7D cell lines carry a mutant p. MCF7 cell line (Bartek et al., 1. None of the new zebrafish monoclonal antibodies recognized human p. Immunoblotting of total protein extracts from these cell lines did reveal that three of the monoclonals (Table 1, column 4) displayed cross- reactivity with other proteins in the cell lines. The band positions of these proteins are not indicative of the human p. All the three cross- reactive monoclonals, namely, ZFp. ZFp. 53- 4. 1 and ZFp. Ig. M isotype. Immunoblots of whole cell extracts of the p. H1. 29. 9 cell line showed that all the monoclonal antibodies were able to detect the p. Table 1, column 5). With the exception of ZFp. H1. 29. 9 cells (Table 1, column 6). Epitope map of the zebrafish p. Functional domains of p. Gen. Bank accession number P7. TA (transactivation domain), DNA- BD (DNA binding domain), BNLS (bipartite nuclear localization signal), OD (oligomerization domain) and BR (basic C- terminal region). Amino acid numbers for both functional domains and antibody epitopes are indicated. The arrows indicate the location of the N terminal amino acid of the mapped epitope. Full figure and legend (1. K)Having established the specificity of the monoclonal antibodies, the ability of the antibodies to detect p. To induce p. 53 protein, 2- day post- fertilization (dpf) zebrafish embryos were treated overnight with 5. Preliminary immunohistochemistry staining showed that monoclonal antibodies ZFp. ZFp. 53- 5. 1, ZFp. ZFp. 53- 9. 1 were able to show strong positive nuclear staining in certain regions of the embryos (Table 1, column 7). Other m. Abs that were negative in the initial immunohistochemistry run were not used further in the antibody characterization studies. Untreated embryos failed to show any immunoreactivity. ZFp. 53- 5. 1 monoclonal antibody as a reagent for zebrafish study in the p. As the monoclonal antibody ZFp. The expression of p. M2. 14. K mutant embryos was examined by immunoblotting. The tp. 53. M2. 14. K mutant line (Berghmans et al., 2. DNA at p. 53- specific consensus sequences. To induce p. 53 protein expression, embryos at 2 dpf were exposed for 1. In a separate experiment, embryos were given a dose of 2. Gy . Immunoblotting results showed that the ZFp. Da in both the wild- type (Figure 3a) and tp. M2. 14. K mutant (Figure 3b) embryo total protein extracts. The p. 53 protein band in the p. H1. 29. 9 cell line (Figure 3a, lane 7) and purified zebrafish p. Figure 3a, lane 8) migrated slower than the p. The intensity of the p. R- roscovitine or irradiated. This indicates that p. To confirm that protein expression could be manipulated at all, one- cell stage wild- type embryos were injected with a p. Figure 3a, lane 2) as compared to untreated embryos (Figure 3a, lane 1), thus confirming the specificity of the antibody in immunoblotting. Detection of p. 53 protein expression by immunoblotting of total protein extracts of 3 dpf zebrafish wild- type embryos and zebrafish tp. M2. 14. K mutant embryos using ZFp. Primary antibody was used at a concentration of 1 . Lanes 6–8 denote controls of untransfected H1. H1. 29. 9 and purified zebrafish p. Lanes 5–7 denote controls of untransfected H1. H1. 29. 9 and purified zebrafish p. Proliferating- cell nuclear antigen antibody was used to verify equal loading amounts of total protein extract of 1. No or low levels of p. Full figure and legend (3. K)Induction of p. R- roscovitine was examined in situ by immunohistochemistry. Zebrafish embryos were treated with 5. Untreated embryos, which had been injected with p. Figure 4a). Embryos injected with a negative control morpholino gave strong nuclear staining in the gut epithelium after the induction of p. R- roscovitine exposure (Figure 4b, inset). The nuclear staining in the intestinal epithelium was not seen when p. R- roscovitine (Figure 4c). This clearly demonstrates that R- roscovitine alters p. ZFp. 53- 5. 1 using immunohistochemistry to stain the tissues in situ. The failure to detect this effect by immunoblotting may be due to the relatively small number of cells responding in the context of the whole animal, so that biochemically the induced protein is highly diluted. Immunohistochemical staining of p. ZFp. 53- 5. 1 hybridoma supernatant on formalin- fixed transverse (a–c) and sagittal (d–g) sections of 3 dpf zebrafish wild- type embryos. Insets show the intensity of nuclear staining in the intestinal epithelium (a) No staining pattern was observed in the gut (gt) for p. R- roscovitine. A moderate amount of staining was also seen in the myotomes. Furthermore, other tissues such as the ganglion cell layer (gcl), cells (rc) of the developing eyes and parts of the developing central nervous system such as telencephalon (tel) and basal plate (bp) also displayed strong nuclear staining indicative of p. Untreated early embryos (2 dpf) do not show any staining (Figure 4d). However, when 2 dpf embryos were exposed overnight to 5. Sagittal sections revealed that the nuclear staining reflecting p. There was also a strong increase of p. Figure 4d). To see the effects of radiation- induced stress versus chemical- induced stress, the expression of p. Similar to the effects of R- roscovitine exposure, embryos exposed to 2. Gy of . Importantly, immunohistochemical examination of tp. M2. 14. K mutant embryos also revealed a dramatic increase in p. Facebook. Entfernen. Wir verwenden Cookies, um Inhalte zu personalisieren, Werbeanzeigen ma. 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