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91.
Schmidt K Marx A de Graaf AA Wiechert W Sahm H Nielsen J Villadsen J 《Biotechnology and bioengineering》1998,58(2-3):254-257
Conventional metabolic flux analysis uses the information gained from determination of measurable fluxes and a steady-state assumption for intracellular metabolites to calculate the metabolic fluxes in a given metabolic network. The determination of intracellular fluxes depends heavily on the correctness of the assumed stoichiometry including the presence of all reactions with a noticeable impact on the model metabolite balances. Determination of fluxes in complex metabolic networks often requires the inclusion of NADH and NADPH balances, which are subject to controversial debate. Transhydrogenation reactions that transfer reduction equivalents from NADH to NADPH or vice versa can usually not be included in the stoichiometric model, because they result in singularities in the stoichiometric matrix. However, it is the NADPH balance that, to a large extent, determines the calculated flux through the pentose phosphate pathway. Hence, wrong assumptions on the presence or activity of transhydrogenation reactions will result in wrong estimations of the intracellular flux distribution. Using 13C tracer experiments and NMR analysis, flux analysis can be performed on the basis of only well established stoichiometric equations and measurements of the labeling state of intracellular metabolites. Neither NADH/NADPH balancing nor assumptions on energy yields need to be included to determine the intracellular fluxes. Because metabolite balancing methods and the use of 13C labeling measurements are two different approaches to the determination of intracellular fluxes, both methods can be used to verify each other or to discuss the origin and significance of deviations in the results. Flux analysis based entirely on metabolite balancing and flux analysis, including labeling information, have been performed independently for a wild-type strain of Aspergillus oryzae producing alpha-amylase. Two different nitrogen sources, NH4+ and NO3-, have been used to investigate the influence of the NADPH requirements on the intracellular flux distribution. The two different approaches to the calculation of fluxes are compared and deviations in the results are discussed. Copyright 1998 John Wiley & Sons, Inc. 相似文献
92.
Site-directed mutagenesis of the simian virus 40 large T-antigen gene: replication-defective amino acid substitution mutants that retain the ability to induce morphological transformation. 总被引:7,自引:7,他引:0 下载免费PDF全文
We used a heteroduplex deletion loop mutagenesis procedure for directing sodium bisulfite-induced mutations to specific sites on viral or plasmid DNA to generate a series of SV40 large T-antigen point mutants. The mutations were directed to a region of the T-antigen gene, 0.5 map units, that is thought to be important for interaction of the protein with the viral origin of DNA replication. Of the 16 mutants reported here, 10 had lost the ability to replicate their DNA, and 3 others showed a reduced level of replication compared to wild type. All of the mutants tested were capable of transforming rat cells in culture by the dense focus assay. We conclude that the sequences of the early region around 0.5 map units are critical for the replication of viral DNA but not for the transformation function of T antigen. 相似文献
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André M Siqueira Lucas I Coutinho Rafael L Gurgel Willian CS Su Luiz M Carvalho Silvana G Benzecry Aline CC Alencar Márcia AA Alexandre Maria Gra?as C Alecrim Marcus VG Lacerda 《Memórias do Instituto Oswaldo Cruz》2014,109(5):540-545
Plasmodium vivax is the most widespread parasite causing malaria, being
especially prevalent in the Americas and Southeast Asia. Children are one of the
most affected populations, especially in highly endemic areas. However, there are
few studies evaluating the therapeutic response of infants with vivax malaria.
This study retrospectively evaluated the parasitaemia clearance in children
diagnosed with vivax malaria during the first five days of exclusive treatment
with chloroquine (CQ). Infants aged less than six months old had a significantly
slower parasitaemia clearance time compared to the group of infants and children
between six months and 12 years old (Kaplan-Meier survival analysis; Wilcoxon
test; p = 0.004). The impaired clearance of parasitaemia in younger children with
vivax malaria is shown for the first time in Latin America. It is speculated that
CQ pharmacokinetics in young children with vivax malaria is distinct, but this
specific population may also allow the detection of CQ-resistant parasites during
follow-up, due to the lack of previous immunity. 相似文献
99.
Uwe Thissen Suzan Wopereis Sjoerd AA van den Berg Ivana Bobeldijk Robert Kleemann Teake Kooistra Ko Willems van Dijk Ben van Ommen Age K Smilde 《BMC bioinformatics》2009,10(1):52
Background
In the fields of life sciences, so-called designed studies are used for studying complex biological systems. The data derived from these studies comply with a study design aimed at generating relevant information while diminishing unwanted variation (noise). Knowledge about the study design can be used to decompose the total data into data blocks that are associated with specific effects. Subsequent statistical analysis can be improved by this decomposition if these are applied on selected combinations of effects. 相似文献100.
Cláudia Rosa‐Ferreira David E Gordon Xavier Franch‐Marro Andrew A Peden Jean‐Paul Vincent Sean Munro 《The EMBO journal》2010,29(2):304-314
Genetic screens in the yeast Saccharomyces cerevisiae have identified many proteins involved in the secretory pathway, most of which have orthologues in higher eukaryotes. To investigate whether there are additional proteins that are required for secretion in metazoans but are absent from yeast, we used genome‐wide RNA interference (RNAi) to look for genes required for secretion of recombinant luciferase from Drosophila S2 cells. This identified two novel components of the secretory pathway that are conserved from humans to plants. Gryzun is distantly related to, but distinct from, the Trs130 subunit of the TRAPP complex but is absent from S. cerevisiae. RNAi of human Gryzun (C4orf41) blocks Golgi exit. Kish is a small membrane protein with a previously uncharacterised orthologue in yeast. The screen also identified Drosophila orthologues of almost 60% of the yeast genes essential for secretion. Given this coverage, the small number of novel components suggests that contrary to previous indications the number of essential core components of the secretory pathway is not much greater in metazoans than in yeasts. 相似文献