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31.
Michio Masuda Tsuyoshi Abe Shinji Sato Teruaki Suzuki Minoru Suzuki 《Journal of phycology》1997,33(2):196-208
Many morphologically similar, but chemically distinct, populations have been found in the marine red alga Laurencia nipponica Yamada (Rhodomelaceae, Ceramiales) growing in Japan. Each chemical type is characterized by a specific end-product of halogenated secondaly metabolite synthesis: chamigrane-type sesquiterpenoids such as prepacifenol and halochamigrene epoxide and C15 bromoethers such as laurencin, laureatin, isoprelaurefucin, epilaurallene, and kumausallene. These seven types of secondary metabolite syntheses remained the same in the wild and under various culture conditions. Because bromoethers and terpenoids are probably synthesized by different metabolic pathways, it is virtually certain that different sets of enzymes participate in their synthesis. Prepacifenol- and laureatin-producing populations were selected as representatives of terpenoid and bromoether groups, respectively. F1 tetrasporophytes derived from crosses between reciprocal, female and male gametophytes of prepacifenol- and laureatin-producing strains bore both types of metabolites, suggesting that the genes Producing these enzyme systems are encoded by nuclear genomes. The F1 gametophytes resulting from the reciprocal crosses produced either prepacifenol or laureatin, and the four individuals derived from spore tetrads (a set of tetraspores derived from a single tetrasporangium) produced either prepacifenol or laureatin in a 1:1 ratio, indicating that genes participating in terpenoid synthsis and those participating in bromoether synthesis are on different loci of homologous chromosomes and are segregated at meiosis (tetrasporogenesis). One individual of this interpopulational F1 gamtophyte produced both parental types of metabolite, perhaps indicating the occurrence of a recombination type. Natural hybrid individuals, including such recombination-type gametophytes, were found in a sympatric locality at which these two chemical types occur. F1 tetrasporophytes derived from crosses between respective prepacifenol- and laureatin-producing strains and their F1 gametohytes produced only parental-type metabolite-producing plants. These results indicate that the diverse chemical types can be referred to as races (chemical races). 相似文献
32.
Shinji Hosoi Mitsuo Satoh Hiromasa Miyaji Tatsunari Nishi Tamio Mizukami Mamoru Hasegawa Seiga Itoh Tatsuya tamaoki 《Cytotechnology》1995,19(1):1-10
Pro-UKS1 was designed as a thrombin-resistant derivative of pro-urokinase (pro-UK) by introducing a glycosylation site using site-directed mutagenesis. An expression plasmid for pro-UKS1, pMo1UKS1SEd1-5, was constructed and introduced into Namalwa KJM-1 cells (Hosoiet al., 1988), and cells resistant to G418 and Methotrexate (MTX) were obtained. Amongst them, the highest pro-UKS1 producer (resistant to 500 nM of MTX), clone 41-8, was selected and further characterized. Clone 41-8 was cultured in serum-free ITPSGF medium (Hosoiet al., 1988). Under the conventional conditions, the concentration of pro-UKS1 reached 26 g ml–1. Addition of glucose and tri-iodothyronine (T3) improved productivity, and the maximal productivity of pro-UKS1 was 67 g ml–1 day–1. In this conditioned medium, content of pro-UKS1 was above 80% of total proteins.Abbreviations BSA
bovine serum albumin
- dhfr
dihydrofolate reductase
- HEPES
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- kb
kilobase pairs
- kDa
kilodaltons
- MTX
Methotrexate
- PBS
phosphate buffered saline
- pro-UK
pro-urokinase
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- T3
tri-iodothyronine
- Tween-PBS
phosphate buffered saline containing 0.05% Tween 80 相似文献
33.
Yoshizawa Itsuo Nagata Kyoko Ohuchi Ryoko Itoh Shinji Kanaiwa Yoshio Amiya Takashi 《Steroids》1980,36(1):87-96
Two D-homosteroids were isolated from the hydrolyzate of 5β-pregnane -3α,20α-diol disulfate (II) when it was refluxed in 3N hydrochloric acid. The structures of these steroids have been elucidated as 17α-methyl-D-homo-5β-androstane-3α, 17aβ-diol (VI) and 17α-methyl-17aγb-chloro-D-homo-5β-androstan-3α-ol (VIII) by instrumental analyses. The former was identical with a synthetic specimen derived from 5β-pregnane-3α,20β-diol di-sulfate (IV) by uranediol rearrangement. The main hydrolyzates obtained were 17α-ethyl-17β-methyl-18-nor-5β-androst-13-en-3α-ol (V) and 5β-pregnane-3α, 20α-diol (III). 相似文献
34.
Fumio Sawada Yasuhiko Miyauchi Hiroshi Tanaka Shinji Matsumoto 《Biochemical and biophysical research communications》1982,104(2):657-663
A novel method for the preparation of intact chromatin from the slime mold which retains the property of RNA synthesis is described. Preparations from G2-cells were highly active, while those from metaphase-cells were inactive. The plasmodial cells were disrupted by gentle homogenization on a polyethylene sieve in a neutral isotonic sucrose medium containing Mg++, deoxycholate and EGTA, a Ca++-chelating agent. The nuclei were lysed in a hypotonic buffer without use of EDTA and chromatin was precipitated by centrifugation after addition of Mg++. 相似文献
35.
Motohiro Morioka† Kohji Fukunaga† Setsuko Yasugawa† Shinji Nagahiro Yukitaka Ushio Eishichi Miyamoto† 《Journal of neurochemistry》1992,58(5):1798-1809
We have investigated regional and temporal alterations in Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) and calcineurin (Ca2+/calmodulin-dependent protein phosphatase) after transient forebrain ischemia. Immunoreactivity and enzyme activity of CaM kinase II decreased in regions CA1 and CA3, and in the dentate gyrus, of the hippocampus early (6-12 h) after ischemia, but the decrease in immunoreactivity gradually recovered over time, except in the CA1 region. Furthermore, the increase in Ca2+/calmodulin-independent activity was detected up to 3 days after ischemia in all regions tested, suggesting that the concentration of intracellular Ca2+ increased. In contrast to CaM kinase II, as immunohistochemistry and regional immunoblot analysis revealed, calcineurin was preserved in the CA1 region until 1.5 days and then lost with the increase in morphological degeneration of neurons. Immunoblot analysis confirmed the findings of the immunohistochemistry. These results suggest that there is a difference between CaM kinase II and calcineurin in regional and temporal loss after ischemia and that imbalance of Ca2+/calmodulin-dependent protein phosphorylation-dephosphorylation may occur. 相似文献
36.
Phenylalanine Ammonia-Lyase Genes from Pisum sativum: Structure, Organ-Specific Expression and Regulation by Fungal Elicitor and Suppressor 总被引:3,自引:0,他引:3
37.
Masayuki Nakamichi Shinji Imakawa Yasuo Kojima Ayuko Natsume 《Primates; journal of primatology》1992,33(3):413-418
Parturition behavior of a multiparous female and her interactions with group members throughout the birth process were recorded
for a free-ranging Japanese monkey (Macaca fuscata). The female showed evidence of 18 contractions during the 35 min prior to delivery, with a mean duration and a mean intercontraction
interval of 30 sec and 96 sec, respectively. These values were similar to those in individually caged Japanese monkeys. Some
adult females remained in proximity to the female who was giving birth during the prepartum phase, and her 2-year-old daughter
watched the delivery of the infant. Even during the prepartum phase the female moved in order to keep up with the group which
traveled from the feeding site to a sleeping site in the forest. 相似文献
38.
Shinji Kakudo Kazumasa Yoshikawa Mikio Tamaki Etsuo Nakamura Hiroshi Teraoka 《Applied microbiology and biotechnology》1992,38(2):226-233
Summary In order to obtain a large quantity of glutamic-acid-specific endopeptidase of Staphylococcus aureus ATCC12600 (SPase) without cultivating its pathogenic host bacterium, expression plasmids enabling secretion of SPase from Bacillus subtilis were constructed by inserting the SPase gene into B. subtilis-Escherichia coli shuttle vectors. B. subtilis harbouring a simple recombinant plasmid containing the coding and the 5-flanking regions of SPase in the shuttle vector pHY300PLK secreted 22 mg/l of SPase into the medium. As this level was lower than that of the natural strain (45 mg/l), we tried to increase the expression level by constructing a series of hybrid plasmids with the following features: (1) the terminator sequence of the alkaline protease gene from B. subtilis, (2) the promoter and the leader sequences of the -amylase gene or of alkaline protease gene from B. amyloliquefaciens, (3) the vector pHY300PLK and the fused vector of pHY300PLK and pUB110. By using a variety of hybrid plasmids, the resulting transformants secreted SPase at levels of 33–120 mg/l. The recombinant SPase isolated from the medium was indistinguishable from the natural one with respect to its behaviour on sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting as well as its enzyme activity.Correspondence to: S. Kakudo 相似文献
39.
The relationship between dissipation of the flash-induced membranepotential across the thylakoid membrane and the high energystate was studied in Zea mays leaves. The dark decay of theflash-induced 515-nm absorbance change was accelerated by shortpreillumination of the leaf. No acceleration of the decay bypreillumination was observed when leaves were incubated in argonor CO2 gas or treated with DCMU. These effects of preilluminationand incubation were reversible. The delayed fluorescence from chlorophyll a was reversibly decreasedby incubating leaves in argon or CO2 gas, though the modes ofdepression were somewhat different from each other. In leavesincubated in argon or CO2 gas, the phase of slow decrease ofthe intensity of prompt fluorescence during illumination reversiblydisappeared. The results suggested that the dissipation of membrane potentialgenerated by a flash was accelerated after the energizationof chloroplasts in leaves, probably by increased H permeabilityof the thylakoid membrane. O2 was important in maintaining (indarkness) and forming (under illumination) the high energy statein chloroplasts in intact leaves. (Received October 1, 1980; Accepted December 15, 1980) 相似文献
40.
Cartilage-derived factor (CDF) : II. Somatomedin-like action on cultured chondrocytes 总被引:2,自引:0,他引:2
Yukio Kato Yoshio Nomura Mitsuko Tsuji Hiroaki Ohmae Masahiko Kinoshita Shinji Hamamoto Fujio Suzuki 《Experimental cell research》1981,132(2):339-347
Previously, we showed that fetal bovine cartilage contains a polypeptide that stimulates the incorporation of [35S]sulfate into proteoglycans synthesized by rat and rabbit costal chondrocytes in culture. In this paper, we report that the cartilage-derived factor (CDF) increases not only [35S]sulfate incorporation but also [3H]thymidine incorporation into rabbit chondrocytes in monolayer culture. The dose-response curve of CDF stimulation of DNA synthesis was similar in profile to that of CDF stimulation of proteoglycan synthesis. In addition, CDF markedly enhanced [3H]uridine incorporation into rabbit chondrocytes and significantly enhanced [3H]serine incorporation into total protein. These findings indicate that fetal bovine cartilage contains a factor that shows somatomedin-like activity in monolayer cultures of rabbit chondrocytes. 相似文献