In addition, we show that base pair mutations in the vRNA promoter can lead to nontemplated dead-end mutations during replication to cRNA in vivo. Based on our observations, we propose C59 wnt cell line a new model for the de novo initiation of influenza virus replication.”
“Accurate characterization of the biochemical pathways of nitric oxide
(NO) is essential for investigations in the field of NO research. To analyze the different reaction pathways of enzymatic and non-enzymatic NO formation, determination of the source of NO is crucial. Measuring NO-related products in biological samples distinguishing between O-14 and (NO)-N-15 offers the opportunity to specifically analyze NO signaling in blood and tissue. The aim of this study was to establish a highly sensitive technique for the specific measurement of NO in an isotopologue-selective manner in biological samples. With the cavity leak-out spectroscopy set-up (CALOS) a differentiation between (NO)-N-14 and (NO)-N-15 is feasible. We describe here the employment of this method for measurements in biological samples. Certified gas mixtures of (NO)-N-14/N-2 and (NO)-N-15/N-2 were used to calibrate the system. click here (NO2-)-N-14 and (NO2-)-N-15 of aqueous and biological samples were reduced in a triiodide solution, and the NO released was detected via CALOS. Gas-phase chemiluminescence
detection (CLD) was used for evaluation. The correlation received for both methods for the detection of NO in the gas-phase was r = 0.999, p < 0.0001. Results obtained using aqueous and biological samples verified that CALOS enables NO measurements with high accuracy (detection limit for (NO2-)-N-14 0.3 pmol and 15 NO2- 0.5 pmol; correlation (NO)-N-14: p < 0.0001, r = 0.975, (NO)-N-15: p < 0.0001, r = 0.969). The CALOS assay represents an extension of NO measurements gmelinol in biological samples, allowing specific investigations of enzymatic and
non-enzymatic NO formation and metabolism in a variety of samples. (c) 2008 Elsevier Inc. All rights reserved.”
“To identify host proteins interacting with Tomato bushy stunt virus (TBSV) replication proteins in a genome-wide scale, we have used a yeast (Saccharomyces cerevisiae) proteome microarray carrying 4,088 purified proteins. This approach led to the identification of 58 yeast proteins that interacted with p33 replication protein. The identified host proteins included protein chaperones, ubiquitin-associated proteins, translation factors, RNA-modifying enzymes, and other proteins with yet-unknown functions. We confirmed that 19 of the identified host proteins bound to p33 in vitro or in a split-ubiquitin-based two-hybrid assay. Further analysis of Cdc34p E2 ubiquitin-conjugating enzyme, which is one of the host proteins interacting with p33, revealed that Cdc34p is a novel component of the purified viral replicase.