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Characterization of Green Tissue-Specific Phytochrome Isolated Immunochemically from Pea Seedlings 总被引:5,自引:0,他引:5
Abe Hiroshi; Yamamoto Kotaro T.; Nagatani Akira; Furuya Masaki 《Plant & cell physiology》1985,26(7):1387-1399
Phytochrome was isolated and purified from light-grown pea (Pisumsativum) seedlings and compared with that from dark-grown seedlingsin terms of spectral and immunochemical properties. Approximately40% of phytochrome in the brushite eluate prepared from light-grownpea tissue bound with a monoclonal anti-pea phytochrome antibody(mAP3), but the remaining 60% did not. Both phytochrome fractionsshowed a typical photoreversible absorbance change after alternatered and far-red actinic irradiations, which was similar to thatof phytochrome from etiolated pea tissue. The peptide mappingof the mAP3-bound phytochrome from light-grown tissue was essentiallythe same as that of the mAP3-bound phytochrome from etiolatedtissue. However, the digestion pattern of the phytochrome thatwas prepared from light-grown tissue but which did not bindto mAP3 was obviously different from that of mAP3-bound phytochrome.Polyclonal anti-pea phytochrome antibodies and mAP5 and 10,however, bound to both the phytochromes. These results suggestthat light-grown tissue contains two phytochrome pools whichare distinct from each other with respect to the primary structureof the phytochrome polypeptide but which share a few commondeterminant sites.
1 Permanent address: Department of Biology, Faculty of Science,Tokyo Metropolitan University, Fukazawa, Tokyo 158, Japan (H.A.), and Department of Botany, Faculty of Science, Universityof Tokyo, Hongo, Tokyo 113, Japan (M. F.). 相似文献
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Chemoattractant-induced tyrosine phosphorylation and activation of microtubule-associated protein kinase in human neutrophils. 总被引:5,自引:0,他引:5
Activation of human neutrophils by the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (fMLP) induces tyrosine phosphorylation of several polypeptides, including a prominent band of approximately 41 kDa. A polypeptide of identical electrophoretic mobility was recognized by a monoclonal antibody raised against a sequence corresponding to amino acids 325-345 of ERK-1, one of a family of mitogen-activated protein (MAP) kinases. To establish the possible identity of these polypeptides, extracts from control and fMLP-treated cells were immunoprecipitated with immobilized antiphosphotyrosine antibodies. Reactivity with anti-ERK-1 antibodies was observed only in the precipitate of chemoattractant-stimulated cells. These data imply that a MAP kinase constitutes at least part of the tyrosine-phosphorylated 41-kDa polypeptide. By using an in vitro renaturation assay, treatment of intact cells with fMLP was found to stimulate several protein kinases, including one of approximately 41 kDa. Renaturation of samples immunoprecipitated with antiphosphotyrosine antibodies revealed the presence of an active protein kinase in chemoattractant-stimulated, but not in control cells. The immunoprecipitated kinase comigrated with the 41-kDa tyrosine phosphorylated polypeptide and the anti-ERK-1 reactive band. We conclude that a MAP kinase closely related or identical to ERK-1 is tyrosine phosphorylated and activated when human neutrophils are stimulated by chemotactic peptides. The rapid phosphorylation of this kinase, which is apparent within seconds, is compatible with a role in the activation of the respiratory burst and/or other neutrophil responses. 相似文献
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H. Furuya Yoh-ji Kukita Sukehisa Nagano Yasuyoshi Sakai Yoriaki Yamashita Hidenao Fukuyama Yuichiro Inatomi Yutaka Saito Ryoko Koike Shoji Tsuji Yasuyuki Fukumaki Kenshi Hayashi Takuro Kobayashi 《Human genetics》1997,100(3-4):450-456
We examined galactosylceramidase (GALC) cDNA in four Japanese patients with adult onset globoid cell leukodystrophy (Krabbe
disease; AO-GLD) by polymerase chain reaction/single-strand conformation polymorphism (PCR-SSCP) analysis, subsequent sequence
determination, and restriction enzyme digestion of PCR products. Initial symptoms were the onset of slowly progressive spastic
paraplegia from the middle of the second decade, and all patients had diminished GALC activity in their leukocytes. We identified
three missense mutations (I66M, G270D, L618S) and one exon-6 skipping (535– 573del). Two of the patients had only the I66M
mutant mRNA, and one only the G270D mutant mRNA. The fourth patient carried a compound heterozygous mutation of 535–573del
and L618S. To determine the enzymatic activities produced by these mutations, we constructed mutated GALC cDNAs and expressed
them in COS-1 cells. Three mutations, viz., G270D, L618S, and exon-6 skipping (535–573del), produced diminished GALC activity
as expected. The I66M mutation in the wild-type GALC cDNA(I289) had normal activity, but when this mutation and the V289 polymorphism
were introduced into the same allele, it had decreased activity. Thus, the combination of a unique mutation and polymorphism
causes conformational change in the GALC enzyme, resulting in low enzymatic activity. AO-GLD mutations, including those found
here, are located in the N-terminus (I66M, G270D, 535–573del) or C-terminus (L618S) of the GALC enzyme, whereas the reported
mutations in the infantile form (IF-GLD) are in the central domain. This difference in mutation sites may affect the clinical
features of GLD.
Received: 4 February 1997 / Accepted: 28 April 1997 相似文献
7.
Tadashi Matsunaga Haruko Takeyama Yuki Miura Takeshi Yamazaki hiroyuki Furuya Koji Sode 《FEMS microbiology letters》1995,133(1-2):137-141
Abstract Screening of fatty acid composition in 150 strains of marine microalgae, cyanobacteria and green algae was carried out, and 20 strains showed relatively high contents of palmitoleic acid. Among them, two cyanobacteria, Phormidium sp. NKBG 041105 and Oscillatoria sp. NKBG 091600, showed an unusually high cis -palmitoleic acid content (54.5% and 54.4% of total fatty acid, respectively). Phormidium sp. NKBG 041105 had the highest cis -palmitoleic acid content per biomass (46.3 mg (g dry cell weight)−1 ), and cis -palrnitoleic acid composition was found to be constant with varying temperature. These results indicate that this cyanobacterium could be considered as a new source for palmitoleic acid. 相似文献
8.
Koh-ichi Enomoto Kishio Furuya Shunichi Yamagishi Takashi Maeno 《Cell biochemistry and function》1993,11(1):55-62
Injection of D -myo-inositol-1,4,5-trisphosphate (IP3) was found to induce a transient increase of intracellular Ca2+ concentration in cancerous mammary cells (MMT060562) and in normal mammary cells treated with epidermal growth factor. Responses to injection of either D -myo-inositol-1,4-bisphosphate (IP2) or D -myo-inositol-1,3,4,5-tetrakisphosphate (IP4) were small or absent. Furthermore, normal mammary cells cultivated with low-protein serum replacement alone or in the presence of differentiation-inducing hormones (insulin + cortisol + prolactin) were less sensitive to IP3. Thapsigargin induced a transient increase of Ca2+ due to the release of Ca2+ from an intracellular pool. There was no difference in the peak heights of the thapsigargin-induced Ca2+ increase when mammary cells were cultivated in the presence or absence of epidermal growth factor or insulin + cortisol + prolactin. These findings suggest that the releasable intracellular Ca2+ pool remained unchanged whereas sensitivity to IP3 increases during the proliferation stage. Mechanical stimulus of a mammary cell induces an increase of intracellular Ca2+ in the stimulated cell. A certain stimulating factor is released from the mechanically stimulated cell into the extracellular space, and it induces an increase of Ca2+ in surrounding cells.18 In contrast, the IP3-induced Ca2+ increase in both cancerous and epidermal growth factor-treated normal mammary cells did not spread to adjacent cells. Therefore, increase of Ca2+ is not sufficient to account for the release of stimulating substances from mammary cells in the mechanically-induced spreading response. 相似文献
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Kanji Takeo Reiko Tanaka Makoto Miyaji Kazuko Nishimura 《FEMS microbiology letters》1995,129(2-3):231-235
Abstract Stationary-phase cells of Cryptococcus neoformans displayed two morphological characteristics: virtually all the cells were unbudded even in the early stationary phase and even when grown in rich media, and average cell size increased from that of exponential-phase cells. DNA contents for small and large stationary-phase cells were determined by quantitative fluorescence microscopy after DNA staining with propidium iodide or DAPI. Small cells contained G, DNA, whereas large unbudded cells had either a G2 or G1 DNA content, indicating that Cr. neoformans can enter into the stationary phase from either the G1 or G2 period. 相似文献
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