Zentrum zur Identifikation von Genfunktionen durch Insertionsmutagenese bei Arabidopsis thaliana (C.V., P.P., A.L., D.L.), and Abteilung fü r Pflanzenzü chtung und Ertragsphysiologie, Max-Planck-Institut fü r Zü chtungsforschung, Carl-von-Linné Weg 10, 50829 Kö ln, Germany (M.T., F.Sc., F.Sa., D.L.); and Institut fü r Biochemie der Pflanzen, Heinrich-Heine-Universität Dü sseldorf, Universitätsstrae 1, 40225 Dü sseldorf, Germany (P.J.)Photosystem I (PSI) of higher plants contains 18 subunits. Using Arabidopsis En insertion lines, we have isolated knockout alleles of the genes psaG, psaH2, and psaK, which code for PSI-G, -H, and -K. In the mutants psak-1 and psag-1.4, complete loss of PSI-K and -G, respectively, was confirmed, whereas the residual H level in psah2-1.4 is due to a second gene encoding PSI-H, psaH1. Double mutants, lacking PSI-G, and also -K, or a fraction of -H, together with the three single mutants were characterized for their growth phenotypes and PSI polypeptide composition. In general, the loss of each subunit has secondary, in some cases additive, effects on the abundance of other PSI polypeptides, such as D, E, H, L, N, and the light-harvesting complex I proteins Lhca2 and 3. In the G-less mutant psag-1.4, the variation in PSI composition suggests that PSI-G stabilizes the PSI-core. Levels of light-harvesting complex I proteins in plants, which lack simultaneously PSI-G and -K, indicate that PSI subunits other than G and K can also bind Lhca2 and 3. In the same single and double mutants, psag-1. 4, psak-1, psah2-1.4, psag-1.4/psah2-1.4, and psag-1.4/psak-1 photosynthetic electron flow and excitation energy quenching were analyzed to address the roles of the various subunits in P700 reduction (mediated by PSI-F and -N) and oxidation (PSI-E), and state transitions (PSI-H). Based on the results, we also suggest for PSI-K a role in state transitions.PSI mediates light-driven electron transport from plastocyanin to ferredoxin across the thylakoid membrane. The PSI complexes in plants and cyanobacteria are basically similar, but there are some notable differences (Scheller et al., 1997): The plant PSI is slightly larger (Kitmitto et al., 1997), trimer formation has been observed only in cyanobacteria (Chitnis and Chitnis, 1993), and the plant PSI core is associated with the light-harvesting complex I (LHCI), which is made up of the four different chlorophyll (Chl)/carotenoidbinding polypeptides Lhca1-4 (Jansson, 1999). In addition, among the 14 subunits that form the core of PSI in flowering plants (PSI-A through -O; Scheller et al., 1997Scheller et al., , 2001; H.V. Scheller, personal communication), four subunits (PSI-G, -H, -N, and -O; Okkels et al., 1989Okkels et al., , 1992 Knoetzel and Simpson, 1993; H.V. Scheller, personal communication) are not present in cyanobacteria. Conversely, no homolog of the cyanobacterial PSI-M has been found yet in higher plants.Most essential for PSI function are the three subunits PSI-A, -B, and -C, which bind the electron acceptors. These acceptor-bi...
The glycoprotein (G-protein) of rabies virus is responsible for viral attachment to the host cell surface and induces virus neutralization antibodies. In the present study, the G-protein gene of rabies virus CVS strain was cloned, sequenced and expressed in the yeast, Pichia pastoris, as a secreted protein, using a simplified DO-stat control feeding strategy. This strategy involves the addition of methanol when the dissolved oxygen (DO) level rises above the setpoint avoiding methanol accumulation and oxygen limitation. The G-protein expression was evaluated by SDS-PAGE, ELISA, and western blot assays. Like native G-protein, the recombinant G-protein was found reactive when it was challenged against specific antibodies. The data indicate that the recombinant G-protein can be easily expressed and isolated, and may be useful as a safe source in the production of diagnostic kits and subunit vaccines to prevent rabies.
The aim of the present study was to compare the endometrial gene expression of epidermal growth factor receptor (EGFR), nodal growth differentiation factor (NODAL), prostaglandin-endoperoxide synthase 2 (PTGS2), oestrogen receptor 1 (ESR1) and progesterone receptor (PGR) in repeat breeder cows (RBC) and non-RBC during diestrus. Endometrial samples were collected by cytobrush technique and stored in RNA stabilizing solution at −20°C until RT-qPCR analysis. Differences in endometrial mRNA expression of selected genes were assessed by ANOVA and simple (r) and the partial correlations (rp) among selected genes were performed. Results demonstrated that mRNA expression of EGFR and NODAL were higher in RBC than in non-RBC (3 and 25-fold change, p < .01 and p < .01, respectively), while the mRNA expression of PTGS2 was lower (1.56-fold change, p < .01). Although there were no differences detected in the mRNA expression of ESR1 and PGR, there was a positive correlation between the expression of ESR1 and EGFR (0.84, p < .05) and a negative correlation between PGR and PTGS2 (−0.49, p < .05). In conclusion, the difference on the endometrial mRNA expression of the genes included in the study between RBC and non-RBC indicates a deregulation of important mechanisms that are vital to establish a successful pregnancy. Thus, the present study provides useful insight as a base for future studies to elucidate the causes of RBC.
La pandemia de la COVID-19 planteó el rápido diseño y aprobación excepcional de diversos métodos de diagnóstico. En la unidad de diagnóstico COVID- 19 de la FCV-UNLP, se realizó el diagnóstico molecular de la presencia de SARS-CoV-2 en 1114 hisopados de pacientes derivados por el Ministerio de Salud de la Provincia de Buenos Aires. Las muestras de ARN fueron purificadas en cabina de seguridad y se analizaron mediante real time RT-PCR con el kit GeneFinder™ para tres targets virales (N, E y RdRp). De 1110 muestras con reacción al control interno, 458 (41,2%) fueron reactivas, 26,9% a tres targets virales, 4,2% a dos y cerca del 10% a uno (principalmente N). El porcentaje de positividad fue similar en el tiempo, aunque la cantidad fue mayor en julio (781 muestras; 315 reactivas) respecto a junio (333 muestras; 143 reactivas). Las muestras de Berisso, Ensenada y La Plata presentaron un porcentaje de positividad significativamente menor al de los demás municipios (27,6% vs. 60,7%). Las muestras de pacientes con tres o más signos relevantes presentaron una mayor positividad (55,6%) y menor reactividad a un único target. Es necesaria la validación interlaboratorios y la estandarización de los métodos para brindar resultados confiables y reproducibles.
Fibropapillomatosis is a debilitating neoplastic disease associated with Chelonid alphaherpesvirus 5 (ChHV5) infection. We detected the Atlantic variant of ChHV5 associated with a fibropapilloma in a green turtle (Chelonia mydas) found stranded on the western coast of Rio de la Plata, Argentina. This is the southernmost registered case for the southwestern Atlantic.
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