全文获取类型
收费全文 | 4738篇 |
免费 | 269篇 |
国内免费 | 3篇 |
出版年
2023年 | 19篇 |
2022年 | 33篇 |
2021年 | 50篇 |
2020年 | 36篇 |
2019年 | 49篇 |
2018年 | 78篇 |
2017年 | 85篇 |
2016年 | 92篇 |
2015年 | 170篇 |
2014年 | 202篇 |
2013年 | 299篇 |
2012年 | 287篇 |
2011年 | 339篇 |
2010年 | 193篇 |
2009年 | 156篇 |
2008年 | 282篇 |
2007年 | 312篇 |
2006年 | 284篇 |
2005年 | 303篇 |
2004年 | 257篇 |
2003年 | 270篇 |
2002年 | 250篇 |
2001年 | 53篇 |
2000年 | 35篇 |
1999年 | 74篇 |
1998年 | 71篇 |
1997年 | 47篇 |
1996年 | 64篇 |
1995年 | 50篇 |
1994年 | 32篇 |
1993年 | 39篇 |
1992年 | 40篇 |
1991年 | 22篇 |
1990年 | 32篇 |
1989年 | 42篇 |
1988年 | 24篇 |
1987年 | 34篇 |
1986年 | 30篇 |
1985年 | 27篇 |
1984年 | 36篇 |
1983年 | 29篇 |
1982年 | 38篇 |
1981年 | 19篇 |
1980年 | 25篇 |
1979年 | 21篇 |
1978年 | 9篇 |
1976年 | 13篇 |
1974年 | 7篇 |
1973年 | 7篇 |
1972年 | 6篇 |
排序方式: 共有5010条查询结果,搜索用时 15 毫秒
61.
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
62.
63.
Cells of Nitella flexilis were made inexcitable by treatmentwith 10 mM KCl for more than 24 h. A Ca2+-sensitive photoproteinaequorin was injected into the cytoplasm of such cells. Forvacuolar per fusion, the central part of an aequorin-loadedcell was immersed in silicone oil, and both cell ends bathedin the perfusion medium were cut off. A large light emissionfrom aequorin was observed when the vacuole was perfused witha hypotonic medium whose osmotic pressure was adjusted to halfof the osmotic pressure of the cell sap. This shows that hydrationof the cytoplasm triggers release of Ca2+ from internal stores,since influx of Ca2+ from silicone oil is excluded. Hydration of cells was induced in another way. Cells were firstdehydrated by transferring them from 10 mM KCl solution to thatwith 250 mM sorbitol added. This procedure did not affect thecytoplasmic streaming. When cells were rehydrated by transferringthem to 10 mM KCl solution, cytoplasmic streaming was eitherstopped or slowed down in a few seconds. A quick light emissionfrom aequorin was observed in the rehydration, evidence thatcytoplasmic streaming was inhibited by an increase in the cytoplasmicCa2+ concentration. Both streaming cessation and aequorin lightemission were observed even in KCl-treated cells which werefurther treated with 5 mM EGTA. Thus, the increase in Ca2+ isconcluded to be caused by the release of Ca2+ from internalstores. These results support our previous hypothesis [Tazawa et al.(1994) Plant Cell Physiol. 35:63] that, in Nitella flexilis,the increase in the concentration of Ca2+ in the cytoplasm whichoccurs on the endoosmotic side of the cell during transcellularosmosis is caused by hydration of the cytoplasm. (Received June 6, 1994; Accepted December 26, 1994) 相似文献
64.
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
65.
Mercurial-sensitive water transport in barley roots 总被引:16,自引:0,他引:16
Masashi Tazawa Eiji Ohkuma Mineo Shibasaka Susumu Nakashima 《Journal of plant research》1997,110(4):435-442
An isolated barley root was partitioned into the apical and basal part across the partition wall of the double-chamber osmometer.
Transroot water movement was induced by subjecting the apical part to a sorbitol solution, while the basal part with the cut
end was in artificial pond water. The rate of transroot osmosis was first low but enhanced by two means, infilitration of
roots by pressurization and repetition of osmosis. Both effects acted additively. The radial hydraulic conductivity (Lpr) was calculated by dividing the initial flow rate with the surface area of the apical part of the root, to which sorbitol
was applied, and the osmotic gradient between the apical and basal part of the root. Lpr which was first 0.02–0.04 pm s−1 Pa−1 increased up to 0.25–0.4 pm s−1 Pa−1 after enhancement. Enhancement is assumed to be caused by an increase of the area of the plasma membrane which is avallable
to osmotic water movement. The increased Lpr is in the same order of magnitude as the hydraulic conductivity (Lp) of epidermal and cortical cells of barley roots obtained
by Steudie and Jeschke (1983). HgCl2, a potent inhibitor of water channels, suppressed Lpr of non-infiltrated and infiltrated roots down to 17% and 8% of control values, respectively. A high sensitivity of Lpr to HgCl2 suggests that water channels constitute the most conductive pathway for osmotic radial water movement in barley roots. 相似文献
66.
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
67.
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
68.
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
69.
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
70.
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