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71.
Construction of a new yeast cloning vector containing autonomous replication sequences from Candida utilis. 总被引:4,自引:1,他引:3
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DNA sequences from the Candida utilis genome which, when cloned into a yeast integration plasmid (YIp5), confer on YIp5 the ability to replicate autonomously in Saccharomyces cerevisiae are described. Several recombinant plasmids which transform S. cerevisiae YNN27 to Ura3+ with an efficiency of 2 X 10(3) transformants per microgram of DNA were obtained. One of the recombinant plasmids, pHMR22 (6.6 kilobases) contains ars (autonomous replication sequence), which is homologous with two different DNA fragments of the C. utilis genome but has no detectable homology to total DNA from Candida albicans, Pachysolen tannophilus, or S. cerevisiae. Restriction and subcloning analyses of pHMR22 showed that Sau3A destroys the functions of cloned ars whereas there are no BamHI, PstI, SalI, HindIII, EcoRI, or PvuII sites in the region of ars which is required for its functional integrity. Thus, pHMR22 appears to be a useful vector for cloning desired genes in S. cerevisiae. 相似文献
72.
Productivity and nutrient uptake of water hyacinth,Eichhornia crassipes I. Effect of nitrogen source
Water hyacinth,Eichhornia crassipes, growth and nutrient uptake rates, as influenced by different N sources and N transformations, were measured using microcosm aquaculture systems. Net productivity was highest in the system receiving equal amounts of NH4 + and NO3 - (at 10 mg N 1-1 each) and decreased in the order of NO3 -, NH4 +, urea (added at 20 mg N 1-1 each), and methane digestor effluent (at 6 mg N 1-1). During the first 7-wk study (average ambient air temperature was 26–28°C), biomass yields were in the range of 19–53 g dry wt m-2 day-1, while between the 8th and 12th wk (average ambient air temperature was 16–22°C), biomass yields were in the range of 10–33 g dry wt m-2 day-1. In the systems with either NH4 + or NO3 -, or both added in equal proportions, about 14–20% of the total yield was contributed by roots, whereas in the system with urea and digestor effluent, roots contributed about 23 and 44% of the total yield, respectively. Nitrogen and P uptake per unit area followed trends similar to biomass yields. Nitrogen uptake rates were in the range of 533–2, 161 mg N m-2 day-1 for the systems receiving NH4 +, NO3 -, and urea, while uptake rates were in the range of 124–602 mg N m-2 day-1 for the system receiving methane digestor effluent. Phosphorus uptake rates were found to be in the range of 59–542 mg P m-2 day-1. Under the most favorable conditions, maximum recorded biomass yield was 53 g dry wt m-2 day-1, with N and P removal rate of 2,161 mg N m-2 day-1 and 542 mg P m-2 day-1, indicating the potential of water hyacinth to produce large amounts of biomass which can be potentially used as a feedstock to produce methane. 相似文献
73.
N-Acylethanolamine phospholipids were produced from endogenous substrates with dog heart mitochondrial and microsomal preparations. With mitochondria the N-acyl group contained 13.8% linoleate, with microsomes only 3.6%. Cardiolipin comprised 18.5% of mitochondrial and 3.3% of microsomal lipid P and contained 93.7 and 72.4% linoleic acid, respectively. Incubation of dog heart subcellular fractions with [1-14C]linoleoyl cardiolipin in the presence of Ca2+ resulted in the formation of N-acylethanolamine phospholipids labeled primarily in the N-acyl and 1-O-acyl moieties. The data indicate that cardiolipin is the major source of linoleic acid used in the N-acylation of ethanolamine phospholipids by transacylase activity. 相似文献
74.
Isolation and functional characterization of hydrocarbon emulsifying and solubilizing factors produced by a Pseudomonas species 总被引:1,自引:0,他引:1
Reddy PG Singh HD Pathak MG Bhagat SD Baruah JN 《Biotechnology and bioengineering》1983,25(2):387-401
Pseudomonas PG-1 cultivated on pristane produced in good amount a heat-stable polymeric substance which showed strong hydrocarbon emulsifying and solubilizing properties. The substance was isolated in crude form and was found to contain 34% protein, 16% carbohydrate, and 40% lipid. The hydrocarbon solubilizing activity of the isolate was strongly inhibited by EDTA but the chelating agent had no effect on the hydrocarbon emulsifying activity. Both activities of the isolate were strongly inhibited by chymotrypsin treatment indicating the importance of the protein moiety for its activity. Hydrocarbon solubilization by the isolate showed a certain degree of specificity to pristane in modest agitation generally used in microbial cultivation, but this specificity was lost by vigorous agitation in a Waring blender. It was proposed that in the first case, solubilization was effected by a solubilizing factor specific to pristane, whereas in the latter case, nonspecific solubilization occurred due to the action of the emulsifying factor. The rate of pristane solubilization by heat-treated culture broth under the conditions of agitation used in cultivation (rotary shaker, 120 rpm) was found to be ca. 750 mg L(-1) h(-1) which was much larger than the maximal pristane uptake rate of 170 mg L(-1) h(-1) observed during microbial growth on the substrate. It was concluded that hydrocarbon solubilization could satisfactorily account for the substrate uptake and growth. 相似文献
75.
Tissue-specific expression of the rat glutathione S-transferases 总被引:9,自引:0,他引:9
Tissue-specific patterns of rat glutathione S-transferase expression have been demonstrated by in vitro translation of purified poly(A) RNAs and by protein purification. Poly(A) RNAs from six rat tissues including heart, kidney, liver, lung, spleen, and testis were used to program in vitro translation with the rabbit reticulocyte lysate system and [35S]methionine. The glutathione S-transferase subunits synthesized in vitro were purified from the translation products by affinity chromatography on S-hexylglutathione-linked Sepharose 6B columns. The affinity bound fractions were analyzed by Na dodecyl SO4-polyacrylamide gel electrophoresis and fluorography. A subunit of Mr = 22,000 detected in the in vitro translation products of poly(A) RNAs from heart, kidney, lung, spleen, and testis is missing from the translation products of liver poly(A) RNAs. This Mr = 22,000 subunit is present only in the anionic glutathione S-transferase fraction purified from rat heart, kidney, lung, spleen, and testis. Purified anionic glutathione S-transferase from rat liver does not contain this subunit. The relative specific activities toward a dozen different substrates also demonstrate the nonidentity between liver and kidney anionic glutathione S-transferases. In addition, among the glutathione S-transferase subunits expressed in the liver, some of them could not be detected in the other tissues investigated. Our results indicate that tissue-specific expression of rat glutathione S-transferases may occur pretranslationally. 相似文献
76.
77.
V Natarajan P C Schmid P V Reddy M L Zuzarte-Augustin H H Schmid 《Journal of neurochemistry》1983,41(5):1303-1312
Abstract: Dog brain homogenates and subcellular preparations incubated in the presence of Ca2+ produced a new phospholipid that was isolated and identified by its infrared spectrum and by chemical degradation as a mixture of 1, 2-diacyl, alkenylacyl, and alkylacyl sn -glycero-3-phospho ( N -acyl)ethanolamines, 50, 45, and 5%, respectively. The N -acyl groups consisted almost exclusively of 16:0, 18:0, and 18:1 fatty acids. Formation of N -acylethanolamine phospholipids from endogenous substrates was linear for about 90 min at approximately 4.5 nmol/h/mg protein and exhibited a pH optimum of 10. Biosynthetic activity was associated with particulate fractions, primarily microsomes, synaptosomes, and mitochondria, but not with myelin. In each case, small amounts (∼0.5 nmol/h/mg protein) of long-chain N -acylethanolamines were also produced. Incubation of dog brain microsomes with 1,2-di[1'-14 C]palmitoyl glycero-phosphocholine yielded N -acylethanolamine phospholipids labeled at both N -acyl (55%) and O -acyl (45%) moieties. It appears that dog brain organelles may contain a phosphatidylethanolamine N -acyl transferase (transacylase) analogous to that recently demonstrated in the myocardial tissue. 相似文献
78.
The Cyt f and P700 contents in leaves of three Sorghum, varietieswere measured, in relation to their carbon assimilation, underdifferent light intensities during growth. At the maximum irradiationused (1,800 µE m2 s1) the ratio of P700to Cyt f was close to unity, whereas under low irradiation (450µE m2 s1) the ratio of P700 to Cyt f rangedfrom two to three. A strikingly positive correlation existedbetween the P700 contents of the leaves and their rates of carbondioxide fixation, dry matter production and Cyt f contents,only when the plants were grown under high light intensities.The P700 content of the leaves in plants grown under low irradiationwas unrelated to the contents of Cyt f. Thus, at a high lightintensity there is a close relationship between the Cyt f andP700 levels, but at low intensities the amounts of electroncarriers and the reaction centre are independent. (Received March 7, 1983; Accepted August 24, 1983) 相似文献
79.
Comparison of AMP and NADH binding to glycogen phosphorylase b 总被引:3,自引:0,他引:3
E A Stura G Zanotti Y S Babu M S Sansom D I Stuart K S Wilson L N Johnson G Van de Werve 《Journal of molecular biology》1983,170(2):529-565
The binding sites for the allosteric activator, AMP, to glycogen phosphorylase b are described in detail utilizing the more precise knowledge of the native structure obtained from crystallographic restrained least-squares refinement than has hitherto been available. Localized conformational changes are seen at the allosteric effector site that include shifts of between 1 and 2 A for residues Tyr75 and Arg309 and very small shifts for the region of residues 42 to 44 from the symmetry-related subunit. Kinetic studies demonstrate that NADH inhibits the AMP activation of glycogen phosphorylase b. Crystallographic binding studies at 3.5 A resolution show that NADH binds to the same sites on the enzyme as AMP, i.e. the allosteric effector site N, which is close to the subunit-subunit interface, and the nucleoside inhibitor site I, which is some 12 A from the catalytic site. The conformations of NADH at the two sites are different but both conformations are "folded" so that the nicotinamide ring is close (approx. 6 A) to the adenine ring. These conformations are compared with those suggested from solution studies and with the extended conformations observed in the single crystal structure of NAD+ and for NAD bound to dehydrogenases. Possible mechanisms for NADH inhibition of phosphorylase activation are discussed. 相似文献
80.
Preparation of oligodeoxyribonucleoside methylphosphonates on a polystyrene support. 总被引:11,自引:8,他引:3
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P S Miller C H Agris A Murakami P M Reddy S A Spitz P O Ts'o 《Nucleic acids research》1983,11(18):6225-6242
An efficient procedure is described for synthesizing deoxyribonucleoside methylphosphonates on polystyrene polymer supports which involves condensing 5'-dimethoxytrityldeoxynucleoside 3'-methylphosphonates. The oligomers are removed from the support and the base protecting groups hydrolyzed by treatment with ethylenediamine in ethanol, which avoids hydrolysis of the methylphosphonate linkages. Two types of oligomers were synthesized: those containing only methylphosphonate linkages, d-Np(Np)nN, and those which terminate with a 5' nucleotide residue, dNp (Np)nN. The latter oligomers can be phosphorylated by polynucleotide kinase, and are separated by polyacrylamide gel electrophoresis according to their chain length. Piperdine randomly cleaves the oligomer methylphosphonate linkages and generates a series of shorter oligomers whose number corresponds to the length of the original oligomer. Apurinic sites introduced by acid treatment spontaneously hydrolyze to give oligomers which terminate with free 3' and 5' OH groups. These reactions may be used to characterize the oligomers. 相似文献