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排序方式: 共有191条查询结果,搜索用时 15 毫秒
41.
A plasmid which can be transferred between Escherichia coli and Pasteurella haemolytica by electroporation and conjugation 总被引:4,自引:0,他引:4
F F Craig J G Coote R Parton J H Freer N J Gilmour 《Journal of general microbiology》1989,135(11):2885-2890
Three broad-host-range plasmids (pRK290, pSa4 and pKT230) and one native Pasteurella haemolytica plasmid (pPH33) were used in transformation experiments with P. haemolytica strains T179 (serotype A1), Y216 (serotype A2) and its capsular-deficient variant Y216/NS1. No transformants were detected with either heat-shock or freeze-thaw techniques. However, by electroporation, all P. haemolytica strains were transformed by pPH33 but not by pRK290 or pSa4. The highest frequency obtained was 91 x 10(4) transformants per microgram of pPH33 DNA with P. haemolytica strain Y216/NS1. Although pPH33 itself was non-transmissible by conjugation, it could be mobilized from Escherichia coli, using the transfer function of the IncP plasmid pRK2013, into P. haemolytica at a frequency of 0.3-2.2 x 10(-3) per recipient cell. 相似文献
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T L Poulos R A Alden S T Freer J J Birktoft J Kraut 《The Journal of biological chemistry》1976,251(4):1097-1103
1. A detailed study of cytochrome C oxidse activity with Keilin-Hartree particles and purified beef heart enzyme, at low ionic strength and low cytochrome C concentrations, showed biphasic kinetics with apparent Km1 = 5 x 10(-8) M, and apparent Km2 = 0.35 to 1.0 x 10(-6) M. Direct binding studies with purified oxidase, phospholipid-containing as well as phospholipid-depleted, demonstrated two sites of interaction of cytochrome c with the enzyme, with KD2 less than or equal to 10(-7) M, and KD2 = 10(-6) M. 2... 相似文献
45.
Stability of xylanases in the presence of methanol and its evaluation on the bleaching capacity 总被引:1,自引:0,他引:1
J. Ruiz R. Angelo J. Freer J. Baeza C. Aguirre E. Curotto N. Durán 《Biotechnology letters》1999,21(4):361-366
A commercial (Cartazyme) and non-commercial (Asperzyme) xylanases were studied. Cartazyme stability in a 0–70% (v/v) methanol at 50°C and 65°C was carried out. No deactivation was found for Cartazyme in the presence of 15% methanol at 50°C. Half-life activity decay (t1/2) of Cartazyme at 50°C in 30%, 50% and 70% methanol solutions were 4.0 h, 2.3 h and 1.2 h, respectively. At 65°C, which is the ozone-alkali-peroxide (ZEP) bleaching temperature, only significant results on Kappa number reduction and selectivity were only observed in 15% methanol (t1/2 30 min) at the Z stage. For the Asperzyme, a t1/2 of 36.5 min at 50°C was found. In the Z stage with Asperzyme in the presence of 25% of methanol, a 20% Kappa number reduction and an improvement of the ZEP sequence of the brightness of 3.1 points were obtained. These results were correlated with the xylanase stability. 相似文献
46.
Production, Purification, and Properties of a Thermostable beta-Glucosidase from a Color Variant Strain of Aureobasidium pullulans 总被引:1,自引:0,他引:1
A color variant strain of Aureobasidium pullulans (NRRL Y-12974) produced beta-glucosidase activity when grown in liquid culture on a variety of carbon sources, such as cellobiose, xylose, arabinose, lactose, sucrose, maltose, glucose, xylitol, xylan, cellulose, starch, and pullulan. An extracellular beta-glucosidase was purified 129-fold to homogeneity from the cell-free culture broth of the organism grown on corn bran. The purification protocol included ammonium sulfate treatment, CM Bio-Gel A agarose column chromatography, and gel filtrations on Bio-Gel A-0.5m and Sephacryl S-200. The beta-glucosidase was a glycoprotein with native molecular weight of 340,000 and was composed of two subunits with molecular weights of about 165,000. The enzyme displayed optimal activity at 75 degrees C and pH 4.5 and had a specific activity of 315 mumol . min . mg of protein under these conditions. The purified beta-glucosidase was active against p-nitrophenyl-beta-d-glucoside, cellobiose, cellotriose, cellotetraose, cellopentaose, cellohexaose, and celloheptaose, with K(m) values of 1.17, 1.00, 0.34, 0.36, 0.64, 0.68, and 1.65 mM, respectively. The enzyme activity was competitively inhibited by glucose (K(i) = 5.65 mM), while fructose, arabinose, galactose, mannose, and xylose (each at 56 mM) and sucrose and lactose (each at 29 mM) were not inhibitory. The enzyme did not require a metal ion for activity, and its activity was not affected by p-chloromercuribenzoate (0.2 mM), EDTA (10 mM), or dithiothreitol (10 mM). Ethanol (7.5%, vol/vol) stimulated the initial enzyme activity by 15%. Glucose production was enhanced by 7.9% when microcrystalline cellulose (2%, wt/vol) was treated for 48 h with a commercial cellulase preparation (5 U/ml) that was supplemented with the purified beta-glucosidase (0.21 U/ml) from A. pullulans. 相似文献
47.
Purification and characterization of the extracellular alpha-amylase from Streptococcus bovis JB1. 总被引:2,自引:2,他引:0 下载免费PDF全文
S N Freer 《Applied microbiology》1993,59(5):1398-1402
The extracellular alpha-amylase (1,4-alpha-D-glucanglucanohydrolase; EC 3.2.1.1) from maltose-grown Streptococcus bovis JB1 was purified to apparent homogeneity by ion-exchange chromatography (Mono Q). The enzyme had an isoelectric point of 4.50 and an apparent molecular mass of 77,000 Da, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was rich in acidic and hydrophobic amino acids. The 15-amino-acid NH2-terminal sequence was 40% homologous with the Bacillus subtilis saccharifying alpha-amylase and 27% homologous with the Clostridium acetobutylicum alpha-amylase. alpha-Amylase activity on soluble starch was optimal at pH 5.0 to 6.0. The enzyme was relatively stable between pH 5.5 and 8.5 and at temperatures below 50 degrees C. When soluble potato starch was used as the substrate, the enzyme had a Km of 0.88 mg.ml-1 and a kcat of 2,510 mumol of reducing sugar.min-1.mg of protein-1. The enzyme exhibited neither pullulanase nor dextranase activity and was 40 to 70% as active on amylopectin as on amylose. The major end products of amylose hydrolysis were maltose, maltotriose, and maltotetraose. 相似文献
48.
Common type 1 fimbriae were isolated from Escherichia coli and their length distribution profile was determined before and after treatment with ultrasound. As fimbriae were shortened, so their haemagglutinating capacity decreased, but their ability to bind to erythrocytes did not decrease to the same extent. Isolated fimbriae did not agglutinate inside-out vesicles prepared from horse erythrocytes or liposomes, suggesting that the binding mechanism was not based on non-specific hydrophobic interactions. The results support a lateral rather than a terminal location for the fimbrial binding site responsible for haemagglutination. 相似文献
49.
The effect of chlordiazepoxide (CDZ) on phenylephrine-induced reflex vagal bradycardia was studied in cats anesthetized with chloralose. The sympathetic component of the reflex was eliminated by either pretreating the animals with propranolol (1 mg/kg, i.v.) or sectioning the spinal cord. In animals pretreated with propranolol, CDZ (3, 10 and 20 mg/kg, i.v.) produced a dose-related inhibition of phenylephrine-induced bradycardia. These doses of CDZ had no significant effect on phenylephrine-induced pressor responses. Similar results were obtained with CDZ in animals with spinal cords transected. Chlordiazepoxide did not prevent bradycardia evoked by electrical stimulation of the peripheral cut-end of the right vagus nerve. These results indicate that CDZ can inhibit reflex vagal bradycardia and that the inhibition involves a central action of the drug. 相似文献
50.
Srinivasabhatt Aswanikumar Barbara Corcoran Elliot Schiffmann Alan R. Day Richard J. Freer Henry J. Showell Elmer L. Becker Candace B. Pert 《Biochemical and biophysical research communications》1977,74(2):810-817
[3H]formylNorleu-Leu-Phe, a potent leucocyte chemoattractant, binds specifically to a site on rabbit neutrophils with an affinity of 1.5 × 10?9 M. Other acylated peptide chemoattractants displace this binding at concentrations closely related to those levels required to elicit chemotaxis. The binding fulfills the major criteria for demonstration of specific receptor sites and indicates that a natural bacterial product and synthetic formyl-peptides produce chemotaxis by the same receptor mechanism. 相似文献