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11.
Deoxyribonucleic Acid Polymerase of Rous Sarcoma Virus: Studies on the Mechanism of Double-Stranded Deoxyribonucleic Acid Synthesis 总被引:7,自引:6,他引:1
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Anthony Faras Lois Fanshier Axel-Claude Garapin Warren Levinson J. Michael Bishop 《Journal of virology》1971,7(5):539-548
The deoxyribonucleic acid (DNA) polymerase of Rous sarcoma virus synthesizes both single- and double-stranded DNA, utilizing the ribonucleic acid (RNA) of the viral genome as the initial template. Results of pulse-chase experiments indicate that the single-stranded DNA serves as unconserved template and precursor for the synthesis of double-stranded DNA. The latter reaction is apparently initiated in association with the viral RNA and may involve a partially double-stranded intermediate form. 相似文献
12.
Linear, Single-Stranded Deoxyribonucleic Acid Isolated from Kilham Rat Virus 总被引:9,自引:9,他引:0
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Kilham rat virus (KRV) was grown in a rat nephroma cell line and was purified by two isopycnic centrifugations in cesium chloride. The virus contains single-stranded deoxyribonucleic acid (DNA) with a molecular weight of approximately 1.6 x 10(6). The DNA was extracted from the virion by both phenol extraction and by 2% sodium dodecyl sulfate at 50 C. KRV DNA, extracted by both procedures, was observed in an electron microscope by using a cytochrome c or diethylaminoethyldextran monolayer. The DNA was also exposed to exonuclease I, an enzyme which hydrolyzes specifically linear, single-stranded DNA. Hydrolysis of 70 to 80% of the DNA was observed. Both the enzymatic and the electron microscope studies support the conclusion that extracted KRV DNA is a single-stranded, linear molecule. The length of the DNA was measured in the electron microscope and determined to be 1.505 +/- 0.206 mum. 相似文献
13.
Surface Antigens of Smooth Brucellae 总被引:29,自引:2,他引:27
Surface antigens of smooth brucellae were extracted by ether-water, phenol-water, trichloroacetic acid, and saline and examined by immunoelectrophoresis and gel diffusion with antisera from infected and immunized rabbits. Ether-water extracts of Brucella melitensis contained a lipopolysaccharide protein component, which was specific for the surface of smooth brucellae and was correlated with the M agglutinogen of Wilson and Miles, a polysaccharide protein component devoid of lipid which was not restricted to the surface of smooth brucellae and was not correlated with the smooth agglutinogen (component 1), and several protein components which were associated with internal antigens of rough and smooth brucellae. Immunoelectrophoretic analysis of ether-water extracts of B. abortus revealed only two components, a lipopolysaccharide protein component, which was correlated with the A agglutinogen, and component 1. Component 1 from B. melitensis and B. abortus showed identity in gel diffusion tests, whereas component M from B. melitensis and component A from B. abortus showed partial identity with unabsorbed antisera and no cross-reactions with monospecific sera. Attempts to prepare monospecific sera directly by immunization of rabbits with cell walls or ether-water extracts were unsuccessful. Absorption of antisera with heavy fraction of ether-water extracts did not always result in monospecific sera. It was concluded (as has been described before) that the A and M antigens are present on a single antigenic complex, in different proportions depending upon the species and biotype, and that this component is a lipopolysaccharide protein complex of high molecular weight that diffuses poorly through agar gel. Components 1, A, and M were also demonstrated in trichloroacetic acid and phenol-water extracts. With all extracts, B. melitensis antigen showed greater diffusibility in agar than B. abortus antigens. After mild acid hydrolysis, B. abortus ether-water extract was able to diffuse more readily. 相似文献
14.
Rat liver microsomes catalyze the hydrolysis of the triphosphates of adenosine, guanosine, uridine, cytidine, and inosine into the corresponding diphosphates and inorganic orthophosphate. The activities are stimulated by Na2S2O4, and inhibited by atebrin, chlorpromazine, sodium azide, and deaminothyroxine. Sodium deoxycholate inhibits the ATPase activity in a progressive manner; the release of orthophosphate from GTP and UTP is stimulated by low, and inhibited by high, concentrations of deoxycholate, and that from CTP and ITP is unaffected by low, and inhibited by high, concentrations of deoxycholate. Subfractionation of microsomes with deoxycholate into ribosomal, membrane, and soluble fractions reveals a concentration of the triphosphatase activity in the membrane fraction. Rat liver microsomes also catalyze the hydrolysis of the diphosphates of the above nucleosides into the corresponding monophosphates and inorganic orthophosphate. Deoxycholate strongly enhances the GDPase, UDPase, and IDPase activities while causing no activation or even inhibition of the ADPase and CDPase activities. The diphosphatase is unaffected by Na2S2O4 and is inhibited by azide and deaminothyroxine but not by atebrin or chlorpromazine. Upon fractionation of the microsomes with deoxycholate, a large part of the GDPase, UDPase, and IDPase activities is recovered in the soluble fraction. Mechanical disruption of the microsomes with an Ultra Turrax Blender both activates and releases the GDPase, UDPase, and IDPase activities, and the former effect occurs more readily than the latter. The GDPase, UDPase, and IDPase activities of the rat liver cell reside almost exclusively in the microsomal fraction, as revealed by comparative assays of the mitochondrial, microsomal, and final supernatant fractions of the homogenate. The microsomes exhibit relatively low nucleoside monophosphatase and inorganic pyrophosphatase activities, and these are unaffected by deoxycholate or mechanical treatment. Different approaches toward the function of the liver microsomal nucleoside tri- and diphosphatases are reported, and the possible physiological role of the two enzymes is discussed. 相似文献
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17.
Lois M. Cherry 《Entomologia Experimentalis et Applicata》1959,2(1):68-76
When larvae of the fly Phormia terraenovae were fed on diets containing fats with different melting points and degrees of saturation, the fat laid down in the depots were effected, though the range of the depot fats was much narrower than that of the fat in the diet. Larvae reared at high temperatures also laid down fat which had a higher melting point and a lower iodine number than did larvae reared at low temperatures.No relation between the properties of the fat and the thermal death point was discovered, though the temperature of rearing had an effect.
Zusammenfassung Larven der Fliege Phormia terraenovae R.-D. wurden im Insektarium bei annähernd 18° C gezogen und mit folgenden Nährstoffen gefüttert: Hefe/Milch, Schweine-, Hammel-, Rindfleisch, Hering.Wenn die Larven völlig erwachsen waren, wurden sie getötet und analysiert. Die Larven wiesen nach allen Ernährungsformen ähnliche Zusammensetzung auf, mit Ausnahme der Jodzahl des Fettkörpers. Diese variierte folgendermaßen: Milch-Hefe-Diät=62, Schwein =70, Hammel=71, Rind=69, Hering=90. Die Unterschiede zwischen mit Schwein, Hammel und Rind ernährten Larven waren nicht signifikant, die anderen Differenzen jedoch stark signifikant.Die Unterschiede zwischen den Jodzahlen der Fette in den verschiedenen Nährstoffen waren größer als diejenigen, die in den mit ihnen gefütterten Larven gefunden wurden (Milch-Hefe=30, Schwein=60, Hering=130).Mit Hammelfleisch bei 35° C ernährte Larven enthielten Fett mit einer Jodzahl von 64 (gegenüber 71 bei den unter 18° C gehaltenen).Der thermale Todespunkt war für alle bei 18° C gezüchteten Larven unabhängig von ihrer Ernährung ungefähr der gleiche. Die bei 35° C gehaltenen Larven wiesen einen annähernd um einen Grad höheren Todespunkt auf.Es scheint also wenig oder gar keine Beziehung zwischen der Zusammensetzung des Fettkörpers der Phormia-Larven und ihrer Resistenz gegen höhere Temperaturen zu bestehen.相似文献
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19.
Dr. Lois Aldwin Robert Toso Robert Goodson Jennie Hunter 《Journal of industrial microbiology & biotechnology》1990,5(4):239-246
Summary The production of streptavidin byStreptomyces avidinii in several different media was examined at 24, 48 and 72 hours. Flask studies indicated that fermentation media containing either complex or multiple carbon sources resulted in higher yields of streptavidin than media with a single carbon source. Streptavidin could be detected in crude fermentation broths by use of a tritiated biotin binding assay. This assay appears to give useful estimates of streptavidin production. Depending upon the medium employed, streptavidin yields ranged from 0.5 mg/l to 53 mg/l. Production was successfully scaled up to ten liter fermentors. Streptavidin was purified in a one step process from centrifuged, concentrated fermentation broths by binding the protein to an iminobiotin column at pH 11 followed by elution at pH 4.0. Recovery percentages varied depending upon the solubility of the fermentation media ingredients. 相似文献
20.
A general assay for restriction endonucleases and other DNA-modifying enzymes with plasmid substrates 总被引:2,自引:0,他引:2
I. Barry Vipond Geoffrey S. Baldwin Mark Oram Symon G. Erskine Lois M. Wentzell Mark D. Szczelkun Timothy J. Nobbs Stephen E. Halford 《Molecular biotechnology》1995,4(3):259-268
A procedure for measuring the activities of enzymes that alter the covalent structure of DNA is described. The assay utilizes
covalently closed circles of DNA as the substrate and yields quantitative data on the fraction of this DNA converted to both
open-circle and linear forms. 相似文献