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41.
42.
Craig J. Coates Catherine L. Turney Marianne Frommer David A. O'Brochta W. D. Warren Peter W. Atkinson 《Molecular & general genetics : MGG》1995,249(2):246-252
Plasmid-based excision assays performed in embryos of two non-drosophilid species using the mariner transposable element from Drosophila mauritiana resulted in empty excision sites identical to those observed after the excision of mariner from D. mauritiana chromosomes. In the presence of the autonomous mariner element Mos1, excision products were recovered from D. melanogaster, D. mauritiana and the blowfly Lucilia cuprina. When a hsp82 heat shock promoter-Mos1 construct was used to supply mariner transposase, excision products were also recovered from the Queensland fruitfly Bactrocera tryoni. Analysis of DNA sequences at empty excision sites led us to hypothesise that the mariner excision/repair process involves the formation of a heteroduplex at the excision breakpoint. The success of these assays suggests that they will provide a valuable tool for assessing the ability of mariner and mariner-like elements to function in non-drosophilid insects and for investigating the basic mechanisms of mariner excision and repair. 相似文献
43.
44.
The state of the art of firefly luciferase research is reviewed with special emphasis on its purification and immobilization. The notion of bioluminescence and its role in APT monitoring is described. The need to purify luciferase and the advantages of immobilization are discussed. An insight into the existing methods of luciferase purification and immobilization is given. The scope of the bioluminescent assay is underlined. 相似文献
45.
An assay is described which detects saxitoxin (STX) and tetrodotoxin (TTX) by their competitive displacement of [3H]saxitoxin from its receptor in rat brain membranes. The assay has a sensitivity of 0.15 ng STX/ml and 0.8 ng TTX/ml for buffer samples. The assay was also applied to detection of these toxins in unextracted human plasma and found to have a sensitivity of 0.5 ng STX/ml and 0.6 ng TTX/ml. The competitive displacement assay appears to be the most sensitive procedure yet for detection of STX and TTX. 相似文献
46.
Purified pyrophosphate: fructose 6-phosphate 1-phosphotransferase (EC 2.7.1.90) was used to measure the inorganic pyrophosphate in unfractionated extracts of tissues of Pisum sativum L. The fructose 1,6-bisphosphate produced by the above enzyme was measured by coupling to NADH oxidation via aldolase (EC 4.1.2.13), triosephosphate isomerase (EC 5.3.1.1) and glycerol-3-phosphate dehydrogenase (EC 1.1.1.8). Amounts of pyrophosphate as low as 1 nmol could be measured. The contents of pyrophosphate in the developing embryo of pea, and in the apical 2 cm of the roots, were appreciable; 9.4 and 8.9 nmol g-1 fresh weight, respectively. The possibility that pyrophosphate acts in vivo as an energy source for pyrophosphate: fructose 6-phosphate 1-phosphotransferase and for UDPglucose pyrophosphorylase (EC 2.7.7.9) is considered. 相似文献
47.
E. B. Sansone A. M. Losikoff W. B. Lebherz III J. A. Poiley 《In vitro cellular & developmental biology. Plant》1981,17(9):811-815
Summary To estimate worker exposures to, and environmental contamination from, test chemicals and organic solvents used in an in vitro
assay to assess the carcinogenic potential of chemicals, sodium fluorescein, a noncarcinogenic fluorescent material, was dissolved
in tissue culture medium used to maintain early passage hamster embryo cells. Personal and environmental samples were taken
over a 14-d period. The assay was performed according to standard procedures in a ventilated glove box or laminar flow safety
cabinet. Considerably more than 99% of the chemical contamination found was recovered from the interiors of the glove box
and hood and from disposable equipment. Contamination outside the containment units (less than 1 μg) resulted from intralaboratory
transport of chemicals, treated cultures, and contaminated equipment. We conclude that the standard operating practices and
procedures provided adequate safeguards for personnel and the environment.
Research sponsored by the National Cancer Institute under Contract N01-CO-75380, with Litton Bionetics, Inc. 相似文献
48.
A procedure for the assay of immobilized tannase with Polyacrylamide gel, collagen and Duolite-S-762 as matrices is described.
It is based on the spectrophotometric determination of gallic acid formed by the enzymatic hydrolysis of tannic acid. The
kinetic parameters of the enzymatic reaction have been studied and an assay procedure has been formulated. This method appears
to be much more accurate than those reported earlier. 相似文献
49.
A centrifugation binding assay has been used to demonstrate the binding of [3H] (±) abscisic acid to membrane-rich fractions prepared from leaves of Vicia faba L. Kinetic analysis of this binding shows evidence of saturation of binding sites with increasing concentration of ligand. Scatchard analysis of these data yields a biphasic plot possibly indicating the presence of two types of binding sites. The dissocation constant for the high affinity site has been calculated to be 3.5×10-8 mol 1-1. 相似文献
50.
Extracts of bovine aorta and nuchal ligament contain several large glycoproteins. The major glycoprotein species has been isolated and has been shown to be collagenase sensitive with an apparent molecular weight of 140,000 daltons. The protein exists in disulphide-bonded aggregates, contains hydroxyproline and hydroxylysine in 1:1 ratio and is unlike any of the known collagen types in amino acid analysis. Its presencein ligament extracts indicates that it is not derived from basement membranes. The evidence suggests that this protein is not derived from the microfibrillar components of the elastic tissues. 相似文献