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101.
The wild-type (dam+) and mutant (damh) forms of the bacteriophage T2 DNA adenine methylase have been partially purified; these enzymes methylate the sequence, 5/t' … G-A-Py … 3′ (Hattman et al., 1978a). However, in vitro methylation studies using phage λ DNA revealed the following: (1) T2 dam+ and damh enzymes differ in their ability to methylate λ DNA; under identical reaction conditions the T2 damh enzyme methylated λ DNA to a higher level than did the dam+ enzyme. However, the respective methylation sites are equally distributed on the l and r strands. (2) Methylation with T2 damh, but not T2 dam+ protected λ against P1 restriction. This was demonstrated by transfection of Escherichia coli (P1) spheroplasts and by cleavage with R·EcoP1. (3) T2 dam+ and damh were similarly capable of methylating G-A-T-C sequences on λ DNA; e.g. λ·dam3 DNA (contains no N6-methyladonine) methylated with either enzyme was made resistant to cleavage by R·DpnII. In contrast, only the T2 damh modified DNA was resistant to further methylation by M·EcoP1 (which methylates the sequence 5′ … A-G-A-C-Py … 3′; Hattman et al., 1978b). (4) λ·dam3 DNA was partially methylated to the same level with T2 dam+ or T2 damh; the two enzymes produced different patterns of G-A-C versus G-A-T methylation. We propose that the T2 dam+ enzyme methylates G-A-C sequences less efficiently than the T2 damh methylase; this property does not entirely account for the large difference in methylation levels produced by the two enzymes. 相似文献
102.
Summary Hyperaccumulators (here defined as species containing at least 500 μg/g dryweight of either copper or cobalt in leaves), are
of interest for the fields of mineral exploration and phytochemistry. Reported hyperaccumulation of copper inAeolanthus biformifolius and the presence of two other species on copper/cobalt mineralization in Shaba (Za?re) led to a survey of these elements
in 49 species of the African genusAeolanthus Mart.A. biformifolius appears to be also a hyperaccumulator of Co (2520 μg/g in leaves, 4300 μg/g in corms). Cobalt levels for most species ofAeolanthus were considerably above typical values for phanerogams and indicate the favorable potential of the genus for further study. 相似文献
103.
The flora of the copper-cobalt ores of Upper Shaba, Zaïre, comprises some 220 taxa including 42 endemics. The origin of this flora is examined and an assessment has been made of its relationship with the flora of the high plateaux steppe-savannah (dilunguan flora). A first group consists of undifferentiated species limited to these two floras. A second group comprises cupriphilous taxa derived from closely-related and widely distributed species observed on the high plateaux and elsewhere. A third group consists of closely-related species or ecotypes confined to the areas. In theSilene burchelli complex (Caryophyllaceae) there is a gradual transition from the widespreadS. burchelli var.angustifolia on the high plateaux to a newly discoveredS. burchelli ecotype from a cupriferous outcrop at Luita, and toS. cobalticola from highly mineralized copper-cobalt deposits at Mindigi. This ecophyletic series provides a gradient of morphological anatomical, and physiological changes whose end members are distinct species. The subject of palaeoendemism and neoendemism in relation to metallophytes of Upper Shaba is also discussed. Colonisation of metalliferous soils by elements of the non-mineralised high plateau is believed to be a neoendemic process. 相似文献
104.
The accuracy of kinetic and stoichiometric data obtained from most laboratory-scale continuous-culture equipment, particularly involving gaseous measurements, may be much lower than many workers realize, despite the use of good quality instruments. For example, errors in specific oxygen uptake measurements (QO(2)) easily can be as high as +/-100%. This article assesses the accuracies of individual instruments and of the overall system in greater detail than has previously been reported and suggestions are made as to how the errors can be reduced to acceptable levels. 相似文献
105.
Citrate and other polyanion binding to ferricytochrome c partially blocks reduction by ascorbate, but at constant ionic strength the citrate-cytochrome c complex remains reducible; reduction by TMPD is unaffected. At a constant high ionic strength citrate inhibits the cytochrome c oxidase reaction competitively with respect to cytochrome c, indicating that ferrocytochrome c also binds citrate, and that the citrateferrocytochrome c complex is rejected by the binding site at high ionic strength. At lower ionic strengths, citrate and other polyanions change the kinetic pattern of ferrocytochrome c oxidation from first-order towards zero-order, indicating preferential binding of the ferric species, followed by its exclusion from the binding site. The turnover at low cytochrome c concentrations is diminished by citrate but not the Km (apparent non-competitive inhibition) or the rate of cytochrome a reduction by bound cytochrome c. Small effects of anions are seen in direct measurements of binding to the primary site on the enzyme, and larger effects upon secondary site binding. It is concluded that anion-cytochrome c complexes may be catalytically competent but that the redox potentials and/or intramolecular behaviour of such complexes may be affected when enzyme-bound. Increasing ionic strength diminishes cytochrome c binding not only by decreasing the ‘association’ rate but also by increasing the ‘dissociation’ rate for bound cytochrome c converting the ‘primary’ (T) site at high salt concentrations into a site similar kinetically to the ‘secondary’ (L) site at low ionic strength. A finite Km of 170 μM at very high ionic strength indicates a ratio of of about 5000. It is proposed that anions either modify the E10 of cytochrome c bound at the primary (T) site or that they perturb an equilibrium between two forms of bound c in favour of a less active form. 相似文献
106.
Free radicals and tissue damage produced by exercise 总被引:11,自引:0,他引:11
K J Davies A T Quintanilha G A Brooks L Packer 《Biochemical and biophysical research communications》1982,107(4):1198-1205
107.
D. E. Brooks 《Molecular reproduction and development》1983,7(4):367-376
Tissue pieces from the caput epididymidis of the rat were incubated in vitro with (35S) methionine to produce radioactive secretory proteins. The radioactive secretory proteins so formed were tested for their ability to bind to washed rat spermatozoa collected from the rete testis and cauda epididymidis, and to rat erythrocytes. The sperm and erythrocytes bound approximately 5% of the total radioactive protein. Binding was protein-specific in that only selected proteins became associated with the cells. Binding was not cell-specific, however, since testicular spermatozoa, caudal spermatozoa, and erythrocytes all bound the same proteins to a similar degree. 相似文献
108.
109.
A recombinant plasmid, designated pUC1002, was constructed by ligation of a HindIII restriction endonuclease fragment of Escherichia coli chromosomal DNA to vector plasmid pMB9. Strains carrying this plasmid were selected by transformation of an E. coli strain bearing the xyl-7 mutation to a xylose-positive (Xyl+) phenotype. Strains containing pUC1002 produced coordinately elevated levels of D-xylose isomerase and D-xylulose kinase. Under appropriate conditions, the isomerase also efficiently catalyzed the conversion of D-glucose to D-fructose. 相似文献
110.
Identification of a negative regulatory region 5' of the human apolipoprotein B promoter 总被引:5,自引:0,他引:5
B Paulweber A R Brooks B P Nagy B Levy-Wilson 《The Journal of biological chemistry》1991,266(32):21956-21961