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11.
Summary The involvement of GATC sites in directing mismatch correction for the elimination of replication errors in Escherichia coli was investigated in vivo by analyzing mutation rates for a gene carried on a series of related plasmids that contain 2, 1 and 0 such sites. This gene encoding chloramphenicol acetyl transferase (Cat protein) was inactivated by a point mutation. In vivo mutations restoring resistance to chloramphenicol were scored in mismatch repair proficient (mut
+) and deficient (mutHLS-) strains. In mut
+ cells, reduction of GATC sites from 2 to 0 increased mutation rates approximately 10-fold. Removal of the GATC site distal to the cat
- mutation increased the rate of mutation less than 2-fold, indicating that mismatch repair can proceed normally with a single site. The mutation rate increased 3-fold after removal of the GATC site proximal to the mutation. In the absence of a GATC site, mutL- and mutS- strains exhibited a 2- to 3-fold increased mutation rate as compared to isogenic mutH- and mut
+ strains. This indicates that 50%–70% of replication errors can be corrected in a mutLS-dependent way in the absence of any GATC site to target mismatch correction to newly synthesized DNA strands. Other strand targeting signals, possibly single strand discontinuities, might be used in mutLS-dependent repair 相似文献
12.
Recently attempts have been made to establish the presence and to determine the metabolic versatility of microorganisms in
the terrestrial deep subsurface at the Savannah River Plant, Aiken, SC, USA. Sediment samples obtained at 20 different depths
of up to 526 m were examined to determine carbon mineralization under aerobic, sulfate-reducing, and methanogenic conditions.
The evolution of14CO2 from radiolabelled glucose was observed under aerobic conditions in all sediments, whereas pyridine was transformed in 50%
of the 20 sediments and indole was metabolized in 85% of the sediments. Glucose mineralization in certain sediments was comparable
to that in the surface environment. Sulfate was reduced in only five sediments, and two were carbon limited. Methane production
was detected in ten sediments amended with formate only after long-term incubations. The transformation of indole and pyridine
was only rarely observed under sulfate-reducing conditions and was never detected in methanogenic incubations. This study
provides information concerning the metabolic capability of both aerobic and anaerobic microorganisms in the deep subsurface
and may prove useful in determining the feasibility of microbial decontamination of such environments. 相似文献
13.
Valérie Maxime Jean-Pierre Pennec Claude Peyraud 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1991,161(6):557-568
Summary The effects of increased ambient salinity (35 mg · ml-1) were studied at 1, 6, and 24 h after direct transfer of rainbow trout from freshwater to seawater. Two series of experiments were carried out successively. The first series was designed to simultaneously study all the respiratory (except Hb affinity for O2), circulatory, and acid-base variables in each fish. In this series, fish were fitted with catheters chronically inserted into the cardiac bulbus, the dorsal aorta, and the opercular and buccal cavities. In the second series, designed to study haemoglobin O2 affinity, fish were fitted with only a dorsal aorta catheter. The ventilatory flow (
) was markedly increased just after transfer (by 55% at 1 h), then more moderately (by 20% at 6 h and 32% at 24 h). The initial hyperventilation peak was associated with frequent couphing motions. These ventilatory changes resulted essentially from increase in ventilatory amplitude. Initially, standard oxygen consumption (MM}O2) decreased slightly, the moderately increased (by 12% at 24 h), so that the oxygen convection requirement (
) increased substantially. In spite of an increased ventilation, the partial pressure of oxygen in arterial blood (P
aO2) decreased slightly at 1 h, prior to returning to control levels, while partial pressure of carbon dioxide in arterial blood (P
aCO2) was not significantly decreased. Gill oxygen transfer factor decreased substantially at 1 h (by 35%) then more moderately (by 7% at 1 h and 12% at 24 h). These results suggest a decrease in gas diffusing capacity of the gills. As P
aCO2 remained approximatively unchanged, the gradual decrease in arterial pH (pHa) from 7.94 to 7.67 at 24 h must therefore be regarded as a metabolic acidosis. The strong ion difference decreased markedly because the concentration of plasma chloride increased more than that of sodium. Arterial O2 content (C
aO2) gradually decreased (by 38% at 24 h) simultaneously with the decrease in pHa, while the ratio P
aO2/C
aO2 increased. In parallel, seawater exposure induced a marked decrease in affinity of haemoglobin for O2, so that at 24 h, P50 was increased by 26% above the value obtained in freshwater-adapted trout. The increase in
could be ascribed initially (at 1 h) to the decrease of P
aO2 and later to a stimulation of respiratory neurons resulting from the lowered medullary interstitial pH. The decrease in C
aO2 could be interpreted mainly as a consequence of a decreased affinity of haemoglobin for O2, likely to be due to the blood acidosis and a predictable increase in chloride concentration within erythrocytes. Cardiac output (
) slightly decreased at 1 h, then progressively increased by 30% at 24 h. Branchial vascular resistance increased at 1 h by 28%, then decreased by 18% of the control value at 24 h. Systemic vascular resistance decreased markedly by 40% at 24 h. As heart rate (HR) remained significantly unchanged, the cardiac stroke volume initially decreased then increased in relation to the changes in
. The increase of
, allowing compensation for the effect of decreased C
aO2 in tissue O2 supply, was interpreted as a passive consequence of the decrease in total vascular resistance occurring during seawater exposure.Abbreviations a.u.
arbitrary units
-
C
aO2
arterial oxygen content
- pH50
arterial pH at P50
-
C
vO2
venous oxygen content
- Hb
haemoglobin
- HR
heart rate
- Hct
hematocrit
- nHill
Hill coefficient
- O2
standard oxygen consumption
-
P
aCO2
arterial partial pressure of carbon dioxide
-
P
aO2
arterial partial pressure of oxygen
-
P
vO2
oxygen partial pressure in mixed venous blood
- P50
oxygen tension at half saturation of haemoglobin
-
P
VA, P
DA
blood pressure in ventral and dorsal aorta
- pHa
arterial pH
-
PIO2, PEO2
oxygen partial pressure of inspired and expired water
-
PO2
oxygen partial pressure
-
cardiac output
- SEM
standard error of mean
- S.I.D.
strong ion difference
- SV
cardiac stroke volume
- TO2
gill oxygen transfer factor
- U
oxygen extraction coefficient
- VA
ventilatory amplitude
- VF
ventilatory frequency
- VRG, VRS
branchial and systemic vascular resistances
-
ventilatory flow
-
ventilatory oxygen convection requirement 相似文献
14.
Jean-Pierre Toutant Jennifer A. Krall Michael K. Richards Terrone L. Rosenberry 《Cellular and molecular neurobiology》1991,11(1):219-230
1. We describe two simple procedures for the rapid identification of certain structural features of glycolipid anchors in acetylcholinesterases (AChEs). 2. Treatment with alkaline hydroxylamine (that cleaves ester-linked acyl chains but not ether-linked alkyl chains) converts molecules possessing a diacylglycerol, but not those with an alkylacylglycerol, into hydrophilic derivatives. AChEs in human and bovine erythrocytes possess an alkylacylglycerol (Roberts et al., J. Biol. Chem. 263:18766-18775, 1988; Biochem. Biophys. Res. Commun. 150:271-277, 1988) and are not converted to hydrophilic dimers by alkaline hydroxylamine. Amphiphilic dimers of AChE from Drosophila, from mouse erythrocytes, and from the human erythroleukaemia cell line K562 also resist the treatment with hydroxylamine and likely possess a terminal alkylacylglycerol. This indicates that the cellular pool of free glycolipids used as precursors of protein anchors is distinct from the pool of membrane phosphatidylinositols (which contain diacylglycerols). 3. Pretreatment with alkaline hydroxylamine is required to render the amphiphilic AChE from human erythrocytes susceptible to digestion by Bacillus thuringiensis phosphatidylinositol-specific phospholipase C (PI-PLC) (Toutant et al., Eur. J. Biochem. 180:503-508, 1989). We show here that this is also the case for the AChE from mouse erythrocytes, which therefore likely possesses an additional acyl chain in the anchor that prevents the action of PI-PLC. 4. In two sublines of K562 cells (48 and 243), we observed that AChE either was directly susceptible to PI-PLC (243) or required a prior deacylation by alkaline hydroxylamine (48). This suggests that glycolipid anchors in AChE of K562-48 cells, but not those in AChE of K562-243 cells, contain the additional acylation demonstrated in AChE from human erythrocytes. These observations illustrate the cell specificity (and the lack of species-specificity) of the structure of glycolipid anchors. 相似文献
15.
Metabolism and Solubilization of Cellulose by Clostridium cellulolyticum H10 总被引:9,自引:2,他引:7 下载免费PDF全文
When Clostridium cellulolyticum was grown with cellulose MN300 as the substrate, the rates of growth and metabolite production were found to be lower than those observed with soluble sugars as the substrate. At low cellulose concentrations, the growth yields were equal to those obtained with cellobiose. The main fermentation products from cellulose and soluble sugars were the same. Up to 15 mM of consumed hexose, a change in the metabolic pathway favoring lactate production similar to that observed with soluble sugars was found to occur concomitantly with a decrease in molar growth yield. With cellulose concentrations above 5 g/liter, accumulation of soluble sugars occurred once growth had ceased. Glucose accounted for 30% of these sugars. A kinetic analysis of cellulose solubilization revealed that cellulolysis by C. cellulolyticum involved three stages whatever cellulose concentration was used. Analysis of these kinetics showed three consecutive enzymatic activity levels having the same Km (0.8 g of cellulose per liter, i.e., 5 mM hexose equivalent) but decreasing values of Vmax. The hypothesis is suggested that each step corresponds to differences in cellulose structure. 相似文献
16.
17.
Summary Alpha-1-microglobulin is a low molecular weight (approximately 30 000 d) glycoprotein present in biological fluids. It is heterogeneous in charge. A monoclonal antibody was used to investigate the tissue distribution of the protein in normal human tissues and cell lines by indirect immunofluorescence and immunoperoxidase techniques. The protein was demonstrable in cells of the monocyte-macrophage lineage, in thymus and T cell dependent areas of spleen, lymph node and tonsils. It was detected in several lymphoid or nonlymphoid cell lines but not in peripheral blood lymphocytes. The microglobulin was also detectable in the cytoplasm of hepatocytes. Finally, it was observed in glandular secretions (sudoral glands and mucosal glands of the digestive tract) where it may be associated with IgA. Possible explanations for the highly divergent results previously reported with polyclonal antisera to 1 microglobulin are discussed. 相似文献
18.
Summary Two populations of Echinochloa crus-galli (Québec, Mississippi) were grown at the Duke University Phytotron under 2 thermoperiods (28°/22°C, 21°/15°C day/night) and 2 CO2 regimes (350 and 675 l l-1). Thermostability, energy of activation (E
a
),K
m (PEP), K
m (Mg++), and specific activity of phospho-enol-pyruvate carboxylase (PEPc) were analyzed in partially purified enzyme preparations of plants grown for 5 weeks. Thermostability of PEPc from extracts (in vitro) and leaves (in situ) was significantly higher in Mississippi plants. In vitro denaturation was not appreciably modified by thermal acclimation but CO2 enrichment elicited higher thermostability of PEPc. In situ thermostability was significantly higher than that of in vitro assays and was higher in Mississippi plants acclimated at 28°/22°C and in plants of the two ecotypes grown at 675 l l-1 CO2. E
a (Q
10 30°/20°C) for PEPc was significantly lower in Québec plants as compared to Mississippi and no acclimatory shifts were observed. Significantly higher K
m's (PEP) in 20°C assays were obtained for Mississippi as compared to Québec plants but values were similar at 30°C and 40°C assays. K
m (Mg++) decreased at higher assay temperatures and were significantly lower for PEPc of the Québec ecotype. No significant changes in K
m (Mg++) values were associated with modifications in temperature on CO2 regimes. PEPc activity measured at 30°C was significantly higher for Québec plants when measured on a leaf fresh weight, leaf area or protein basis but not on a chlorophyll basis. Significantly higher PEPc activity for both genotypes was observed for plants acclimated at 21°/15°C or grown at 675 l l-1 CO2. Net photosynthesis (Ps) and net assimilation rates (NAR) were higher in Québec plants and were enhanced by CO2 enrichment. NAR was higher in plants acclimated at low temperature, while an opposite trend was observed for Ps. PEPc activities were always in excess of the amounts required to support observed rates of CO2 assimilation. 相似文献
19.
Rice root glutamate synthase activity was assayed with various reducing systems. Ferredoxin-dependent glutamate synthase (EC 1.4.7.1) and pyridine nucleotide-dependent glutamate synthase (NADH, EC 1.4.1.14; or NADPH, EC 1.4.1.13) exhibited a strict specificity for the electron donor. The ferredoxin-dependent glutamate synthase from rice roots could accept electrons from photoreduced ferredoxin in an illuminated reconstituted spinach chloroplast system. Thioredoxin, a potent electron carrier, was not able to provide either ferredoxin-dependent or pyridine nucleotide-dependent glutamate synthase with electrons as no glutamate formation was detected in the presence of reduced thioredoxin f or m. 相似文献
20.