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71.
72.
Transport of chimeric proteins that contain a carboxy-terminal targeting signal into plant microbodies 总被引:6,自引:2,他引:4
Makoto Hayashi Masahiro Aoki Akira Kato Maki Kondo Mikio Nishimura 《The Plant journal : for cell and molecular biology》1996,10(2):225-234
Malate synthase is a glyoxysome-specific enzyme. The carboxy-terminal tripeptide of the enzyme is Ser—Arg—Leu (SRL), which is known to function as a peroxisomal targeting signal in mammalian cells. To analyze the function of the carboxy-terminal amino acids of pumpkin malate synthase in plant cells, a chimeric gene was constructed that encoded a fusion protein which consisted of β-glucuronidase and the carboxyl terminus of the enzyme. The fusion protein was expressed and accumulated in transgenic Arabidopsis that had been transformed with the chimeric gene. Immunocytochemical analysis of the transgenic plants revealed that the carboxy-terminal five amino acids of pumpkin malate synthase were sufficient for transport of the fusion protein into glyoxysomes in etiolated cotyledons, into leaf peroxisomes in green cotyledons and in mature leaves, and into unspecialized microbodies in roots, although the fusion protein was no longer transported into microbodies when SRL at the carboxyl terminus was deleted. Transport of proteins into glyoxysomes and leaf peroxisomes was also observed when the carboxy-terminal amino acids of the fusion protein were changed from SRL to SKL, SRM, ARL or PRL. The results suggest that tripeprides with S, A or P at the −3 position, K or R at the −2 position, and L or M at the carboxyl terminal position can function as a targeting signal for three kinds of plant microbody. 相似文献
73.
74.
Successful expression in pollen of various plant species of in vitro synthesized mRNA introduced by particle bombardment 总被引:5,自引:0,他引:5
Toshinori Tanaka Masahiro Nishihara Motoaki Seki Atsushi Sakamoto Kunisuke Tanaka Kohei Irifune Hiromichi Morikawa 《Plant molecular biology》1995,28(2):337-341
Gold particles coated with -glucuronidase (GUS) mRNA with a 5 cap structure that had been synthesized in vitro were introduced, by use of a pneumatic particle gun, into pollen grains of lily (Lilium longiflorum), freesia (Freesia refracta) and tulip (Tulipa gesneriana). A fluorometric assay for the GUS activity indicated that in vitro synthesized GUS mRNA introduced into these pollen cells by particle bombardment was successfully expressed. GUS activity in extracts of the bombarded lily pollen became detectable fluorometrically within 30 min after bombardment, peaked at 6 h, then gradually decreased. This activity changed as a function of the developmental stage of the pollen cell of lily. 相似文献
75.
Nishijima K Hisatsune T Kato H Kohyama M Kakehi M Hachimura S Kaminogawa S 《Cytotechnology》1997,25(1-3):89-100
Feeding of a whole casein diet, which abolished the αs1-casein-specific proliferation and IFN-γ productivity of CD4+ T cells, did not affect the proliferative response of CD8+ T cells with regard to the antigen dose response, cell dose response, kinetics of the proliferation and epitope specificity,
as well as IFN-γ production. To assess the characteristics of the CD8+ T cells, we established αs1-casein-specific CD8+ T cell clones from both casein-fed and control mice. The established clones produced different amount of IFN-γ and IL-10,
and one clone derived from the casein-fed mice produced a remarkable amount of IL-10. The clones from casein-fed mice produced
considerable amounts of TGF-β, while those from control mice produced only small amounts. The possible role of CD8+ T cells in oral tolerance is discussed.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
76.
Akira Uchimura Toshiyuki Shimizu Masahiro Morita Hitomi Ueno Kazuhiro Motoki Hideaki Fukushima Takenori Natori Yasuhiko Koezuka 《Bioorganic & medicinal chemistry》1997,5(12):2245-2249
We compared the immunostimulatory effects of chemically synthesized α-galactosylceramides (α-GalCers), α-glucosylceramides (α-GluCers), 6″-monoglycosylated α-GalCer and 6″- or 4″-monoglycosylated α-GluCer and made the following observations: (1) the length of the fatty acid side chain in the ceramide portions greatly affects the immunostimulatory effects of α-GalCers and α-GluCers; (2) the configuration of the 4″-hydroxyl group of the inner pyranose moiety plays an important role in the immunostimulatory effects of monoglycosylated α-
-pyranosylceramides; (3) the free 4″-hydroxyl group of the inner pyranose of monoglycosylated α-
-pyranosylceramides plays a more important role in their immunostimulatory effects than the free 6″-hydroxyl group. 相似文献
77.
Souichi Satoh †Tatsuo Kimura †Masahiro Toda †Mutuko Maekawa †Satoshi Ono †Hirokazu Narita Hiroyuki Miyazaki Toshihiko Murayama Yasuyuki Nomura 《Journal of neurochemistry》1997,69(5):2197-2205
Abstract: Nitrogen oxides, such as nitric oxide, have been shown to regulate neuronal functions, including neurotransmitter release. We investigated the effect of S-nitroso-l -cysteine (SNC) on noradrenaline (NA) release in the rat hippocampus in vivo and in vitro. SNC stimulated [3H]NA release from prelabeled hippocampal slices in a dose-dependent manner. SNC stimulated endogenous NA release within 30 min to almost five times the basal level in vivo (microdialysis in freely moving rats). In a Na+-containing Tyrode's buffer, SNC-stimulated [3H]NA release was inhibited 30% by the coaddition of l -leucine. In the Na+-free, choline-containing buffer, SNC-stimulated [3H]NA release, which was similar to that in the Na+-containing buffer, was inhibited markedly by l -leucine, l -alanine, l -methionine, l -phenylalanine, and l -tyrosine. The effects of the other amino acids examined were smaller or very limited. The effect of l -leucine was stronger than that of d -leucine. A specific inhibitor of the L-type amino acid transporter, 2-aminobicyclo[2.2.1]-heptane-2-carboxylate (BCH), inhibited the effects of SNC on [3H]NA release in the Na+-free buffer. Uptake of l -[3H]leucine into the slices in the Na+-free buffer was inhibited by SNC, BCH, and l -phenylalanine, but not by l -lysine. The effect of SNC on cyclic GMP accumulation was not inhibited by l -leucine, although SNC stimulated cyclic GMP accumulation at concentrations up to 25 µM, much less than the concentration that stimulates NA release. These findings suggest that SNC is incorporated into rat hippocampus via the L-type-like amino acid transporter, at least in Na+-free conditions, and that SNC stimulates NA release in vivo and in vitro in a cyclic GMP-independent manner. 相似文献
78.
Masahiro Sakaguchi Sakae Inouye Yuichi Takahashi Susumu Kutagiri Hiroshi Yasueda 《Aerobiologia》1995,11(4):265-268
We examined the kinetics of airborne levels of mite allergen particles in a house by combined use of an indoor Burkard air
sampler and immunoblotting. Airborne mite allergens collected on the Burkard sampling tape were transferred onto a nitrocellulose
membrane, reacted with mouse monoclonal anti-mite allergen (Der pI) antibody, then treated with alkaline phosphatase conjugated anti-mouse IgG. Finally, the blotted allergen on the membrane
was reacted with BCIP/NBT phosphatase, and purple spots visible by the naked eye were produced. The shape of the spots was
observed under a microscope, and the spot area was measured by an image processor. This technique might be useful for analyzing
the behavior of airborne allergen particles in indoor environments. 相似文献
79.
Minoru Yonezawa Masahiro Takahata Naoko Banzawa Nobuyuki Matsubara Yasuo Watanabe Hirokazu Narita 《Microbiology and immunology》1995,39(4):243-247
Artificial mutations of Gyrase A protein (GyrA) in Escherichia coli by site-directed mutagenesis were generated to analyze quinolone-resistant mechanisms. By genetic analysis of gyrA genes in a gyrA temperature sensitive (Ts) background, exchange of Ser at the NH2-terminal 83rd position of GyrA to Trp, Leu, Phe, Tyr, Ala, Val, and Ile caused bacterial resistance to the quinolones, while exchange to Gly, Asn, Lys, Arg and Asp did not confer resistance. These results indicate that it is the most important for the 83rd amino acid residue to be hydrophobic in expressing the phenotype of resistance to the quinolones. These findings also suggest that the hydroxyl group of Ser would not play a major role in the quinolone-gyrase interaction and Ser83 would not interact directly with other amino acid residues. 相似文献
80.
Masahiro Goto Hiroshi Sumura Kojiro Abe Fumiyuki Nakashio 《Biotechnology Techniques》1995,9(2):101-104
Summary A novel preparation method for surfactant-coated enzymes has been developed using a W/O emulsion. The enzymatic activity of chymotrypsin in isooctane significantly increased with the coating of surfactants. The surfactant-coated chymotrypsin showed a high enzymatic activity for amidation, although powdered chymotrypsin did not show the activity. Further, the coated enzyme showed a remarkably high storage stability. 相似文献