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81.
Aromatic amino acid auxotrophs of the methanol-utilizing yeast Hansenula polymorpha were effectively selected by the use of nystatin and a medium that inhibits the growth of tyrosine auxotrophs. The procedure resulted in a frequency of aromatic auxotrophs of 2% of survivors and an enrichment of 20-fold. The new procedure also takes less time than traditional procedures. Of the auxotrophic mutants isolated, two-thirds required tyrosine and the remainder were tyrosine-phenylalanine double auxotrophs. 相似文献
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A partially purified D-xylose isomerase has been isolated from cells of Streptomyces albus NRRL 5778 and some of its properties have been determined. D-Glucose, D-xylose, D-ribose, L-arabinose, and L-rhamnose served as substrates for the enzyme with respective Km values of 86, 93, 350, 153, and 312 mM and Vmax values measuring 1.23, 2.9, 2.63, 0.153, and 0.048 mumol min per mg of protein. The hexose D-allose was also isomerized. The enzyme was strongly activated by 1.0 mM Mg2+ but only partially activated by 1.0 mM Co2+. The respective Km values for Mg2+ and Co2+ were 0.3 and 0.003 mM. Mg2+ and Co2+ appear to have separate binding sites on the isomerase. These cations also protect the enzyme from thermal denaturation and from D-sorbitol inhibition. The optimum temperature for ketose formation was 70 to 80 C at pH values ranging from 7 to 9. D-Sorbitol acts as a competitive inhibitor with a Ki of 5.5 mM against D-glucose, D-xylose, and D-ribose. Induction experiments, Mg2+ activation, and D-sorbitol inhibition indicated that a single enzyme (D-xylose isomerase) was responsible for the isomerization of the pentoses, methyl pentose, and glucose. 相似文献
84.
Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. 总被引:225,自引:28,他引:197
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Agarose gel electrophoresis may be employed effectively for the detection and preliminary characterization of plasmid deoxyribonucleic acid (DNA) present in clinical isolates and laboratory strains of gram-negative microorganisms. The method is sensitive and does not require radioisotopes or ultracentrifugation. The estimation of plasmid mass from the extent of DNA migration in gels compares favorably with results obtained by electron microscopy of plasmid DNA purified by equilibrium density centrifugation. The method has proved to be a useful tool for survey work and the epidemiological investigation of plasmid dissemination, as well as an important adjunct to the genetic analysis of plasmids. 相似文献
85.
Summary It was found that the increase of the hydraulic retention time improved the organic matter removal and the conversion of organic to ammonia nitrogen. Methane concentration in the biogas was not affected by the hydraulic retention time.The reactor in operation during one year was not clogged. 相似文献
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87.
James V. McCann Steven R. Bischoff Yu Zhang Dale O. Cowley Veronica Sanchez‐Gonzalez George D. Daaboul Andrew C. Dudley 《Genesis (New York, N.Y. : 2000)》2020,58(7)
Extracellular vesicles (EVs) are abundant, lipid‐enclosed vectors that contain nucleic acids and proteins, they can be secreted from donor cells and freely circulate, and they can be engulfed by recipient cells thus enabling systemic communication between heterotypic cell types. However, genetic tools for labeling, isolating, and auditing cell type‐specific EVs in vivo, without prior in vitro manipulation, are lacking. We have used CRISPR‐Cas9‐mediated genome editing to generate mice bearing a CD63‐emGFPloxP/stop/loxP knock‐in cassette that enables the specific labeling of circulating CD63+ vesicles from any cell type when crossed with lineage‐specific Cre recombinase driver mice. As proof‐of‐principle, we have crossed these mice with Cdh5‐CreERT2 mice to generate CD63emGFP+ vasculature. Using these mice, we show that developing vasculature is marked with emerald GFP (emGFP) following tamoxifen administration to pregnant females. In adult mice, quiescent vasculature and angiogenic vasculature (in tumors) is also marked with emGFP. Moreover, whole plasma‐purified EVs contain a subpopulation of emGFP+ vesicles that are derived from the endothelium, co‐express additional EV (e.g., CD9 and CD81) and endothelial cell (e.g., CD105) markers, and they harbor specific miRNAs (e.g., miR‐126, miR‐30c, and miR‐125b). This new mouse strain should be a useful genetic tool for generating cell type‐specific, CD63+ EVs that freely circulate in serum and can subsequently be isolated and characterized using standard methodologies. 相似文献
88.
Susana Ruiz‐Ruiz Sergio Sanchez‐Carrillo Sergio Ciordia María C. Mena Celia Mndez‐García David Rojo Rafael Bargiela Elisa Zubeldia‐Varela Mnica Martínez‐Martínez Coral Barbas Manuel Ferrer Andrs Moya 《Aging cell》2020,19(1)
Composition of the gut microbiota changes during ageing, but questions remain about whether age is also associated with deficits in microbiome function and whether these changes occur sharply or progressively. The ability to define these deficits in populations of different ages may help determine a chronological age threshold at which deficits occur and subsequently identify innovative dietary strategies for active and healthy ageing. Here, active gut microbiota and associated metabolic functions were evaluated using shotgun proteomics in three well‐defined age groups consisting of 30 healthy volunteers, namely, ten infants, ten adults and ten elderly individuals. Samples from each volunteer at intervals of up to 6 months (n = 83 samples) were used for validation. Ageing gradually increases the diversity of gut bacteria that actively synthesize proteins, that is by 1.4‐fold from infants to elderly individuals. An analysis of functional deficits consistently identifies a relationship between tryptophan and indole metabolism and ageing (p < 2.8e?8). Indeed, the synthesis of proteins involved in tryptophan and indole production and the faecal concentrations of these metabolites are directly correlated (r2 > .987) and progressively decrease with age (r2 > .948). An age threshold for a 50% decrease is observed ca. 11–31 years old, and a greater than 90% reduction is observed from the ages of 34–54 years. Based on recent investigations linking tryptophan with abundance of indole and other “healthy” longevity molecules and on the results from this small cohort study, dietary interventions aimed at manipulating tryptophan deficits since a relatively “young” age of 34 and, particularly, in the elderly are recommended. 相似文献
89.
Jiangong Liu Yulun Zhou Alex Valach Robert Shortt Kuno Kasak Camilo Rey‐Sanchez Kyle S. Hemes Dennis Baldocchi Derrick Y. F. Lai 《Global Change Biology》2020,26(9):4998-5016
The role of coastal mangrove wetlands in sequestering atmospheric carbon dioxide (CO2) and mitigating climate change has received increasing attention in recent years. While recent studies have shown that methane (CH4) emissions can potentially offset the carbon burial rates in low‐salinity coastal wetlands, there is hitherto a paucity of direct and year‐round measurements of ecosystem‐scale CH4 flux (FCH4) from mangrove ecosystems. In this study, we examined the temporal variations and biophysical drivers of ecosystem‐scale FCH4 in a subtropical estuarine mangrove wetland based on 3 years of eddy covariance measurements. Our results showed that daily mangrove FCH4 reached a peak of over 0.1 g CH4‐C m?2 day?1 during the summertime owing to a combination of high temperature and low salinity, while the wintertime FCH4 was negligible. In this mangrove, the mean annual CH4 emission was 11.7 ± 0.4 g CH4‐C m–2 year?1 while the annual net ecosystem CO2 exchange ranged between ?891 and ?690 g CO2‐C m?2 year?1, indicating a net cooling effect on climate over decadal to centurial timescales. Meanwhile, we showed that mangrove FCH4 could offset the negative radiative forcing caused by CO2 uptake by 52% and 24% over a time horizon of 20 and 100 years, respectively, based on the corresponding sustained‐flux global warming potentials. Moreover, we found that 87% and 69% of the total variance of daily FCH4 could be explained by the random forest machine learning algorithm and traditional linear regression model, respectively, with soil temperature and salinity being the most dominant controls. This study was the first of its kind to characterize ecosystem‐scale FCH4 in a mangrove wetland with long‐term eddy covariance measurements. Our findings implied that future environmental changes such as climate warming and increasing river discharge might increase CH4 emissions and hence reduce the net radiative cooling effect of estuarine mangrove forests. 相似文献
90.