全文获取类型
收费全文 | 3628篇 |
免费 | 160篇 |
国内免费 | 193篇 |
出版年
2023年 | 29篇 |
2022年 | 33篇 |
2021年 | 58篇 |
2020年 | 67篇 |
2019年 | 94篇 |
2018年 | 90篇 |
2017年 | 60篇 |
2016年 | 66篇 |
2015年 | 76篇 |
2014年 | 209篇 |
2013年 | 265篇 |
2012年 | 216篇 |
2011年 | 246篇 |
2010年 | 179篇 |
2009年 | 148篇 |
2008年 | 169篇 |
2007年 | 160篇 |
2006年 | 151篇 |
2005年 | 119篇 |
2004年 | 138篇 |
2003年 | 110篇 |
2002年 | 106篇 |
2001年 | 73篇 |
2000年 | 72篇 |
1999年 | 56篇 |
1998年 | 51篇 |
1997年 | 37篇 |
1996年 | 37篇 |
1995年 | 43篇 |
1994年 | 58篇 |
1993年 | 40篇 |
1992年 | 44篇 |
1991年 | 52篇 |
1990年 | 33篇 |
1989年 | 38篇 |
1988年 | 31篇 |
1987年 | 31篇 |
1986年 | 33篇 |
1985年 | 37篇 |
1984年 | 65篇 |
1983年 | 41篇 |
1982年 | 60篇 |
1981年 | 55篇 |
1980年 | 42篇 |
1979年 | 49篇 |
1978年 | 32篇 |
1977年 | 29篇 |
1976年 | 17篇 |
1975年 | 12篇 |
1973年 | 11篇 |
排序方式: 共有3981条查询结果,搜索用时 125 毫秒
101.
以感染肾综合征出血热病毒(HFRSV)的Vero E6细胞为材料,用免疫亲和层析结合制备聚丙烯酰胺凝胶电泳(PAGE)从感染细胞中提纯了HFRSV两种糖蛋白。先用免疫亲和层析从感染细胞的粗制抗原中获得含有四种蛋白的混合液,用[~3H]-氨基葡萄糖在感染细胞中标记病毒糖蛋白,观察到[~3H]-氨基葡萄糖只结合入78K和57K的病毒蛋白。再用制备SDS-PAGE从HFRSV混合液中提纯78K和57K两种蛋白。实验证明这两种糖蛋白均具中和抗原决定簇,57K的糖蛋白尚具血凝活性,初步鉴定表明这两种糖蛋白相当于文献报道的HFRSV G_1和G_2。 相似文献
102.
The structure of the vacuolar ATPase from mesophyll tonoplasts of Mesembryanthemum crystallinum has been studied by electron microscopy using negatively stained specimens of membrane-bound and detergent-solubilized ATPase molecules. We observed a high density of particles on the surface of tonoplast vesicles and “head and stalk” structures on the edge of the membrane, similar to the F0F1-ATPases of mitochondrial and chloroplast membranes. The staining conditions, which are often critical for such small objects, were improved by using methylamine tungstate as negative stain for the membrane-bound ATPase. Compared to other staining solutions generally applied, dissociation of the F1-like enzyme complex from the membrane was best prevented and structural damage of the vesicles was least observed with methylamine tungstate. In freeze-fracture electron microscopy of tonoplast vesicles, where dissociation never occurs since no detergent is used, we also observed “head and stalk” structures on the edge of the membranes, beside many particles on the fracture faces. The detergent-solubilized ATPase forms string-like structures, caused by the aggregation of the hydrophobic membrane-embedded F0-like part of the enzyme. After negative staining the F1-like enzyme complex is arranged alternately along both sides of the string and connected by a narrow stalk. 相似文献
103.
On-line fluorescence-monitoring of the methanogenic fermentation 总被引:2,自引:0,他引:2
On-line in situ fluorescence measurements of the methanogenic fermentation were conducted with reactors receiving either glucose or a mixture of volatile fatty acids as the substrate. The reactors were perturbed from steady-state conditions in order to assess the response of fluorescencemonitoring probes. Two fluorescence-monitoring probes were evaluated over a period of 8 months; they performed in a consistent manner, and their response was not significantly affected by the changes in pH and redox potential encountered during routine reactor operation. A commercially available probe, designed to measure NAD(P)H, demonstrated particular promise for detecting imbalance caused by the entry of air, inhibitor addition and was capable of distinguishing between different substrates. This fluorescence-monitoring probe detected imbalance more rapidly than other on-line measurements such as pH, Eh, or gas production, or off-line measurements such as volatile fatty acid concentration or gas composition. An experimental fluorescence-monitoring probe, designed to measure coenzyme F(420), also showed some promise in this regard. The response of the fluorescence-monitoring probes also revealed details of the metabolic routes in the reactors and the probes represent a useful research tool. For example, a failure to observe the characteristic response of the NAD(P)H-monitoring probe to formate addition during the metabolism of acetate, propionate, or glucose strongly suggests that any formate liberated during their catabolism is degraded via a different route to exogenously added formate. 相似文献
104.
Eijiro Watanabe Masaaki Wachi Makari Yamasaki Kazuo Nagai 《Molecular & general genetics : MGG》1992,234(3):346-352
Summary The SopA, B, C genes of the F plasmid play an essential role in plasmid partitioning during cell division in Escherichia coli. In this paper, the products of the sopA and sopB genes were isolated and their biochemical activities studied. [-32P]ATP was cross-linked to the SopA protein by UV irradiation; this cross-linking was observed only in the presence of magnesium ion, and was competitively inhibited in the presence of non-radioactive ATP, ADP and dATP, but not other NTPs or dNTPs. In contrast, no ATP binding activity was detected for the SopB protein. The SopA protein showed a modest magnesium ion-dependent ATPase activity and this activity was stimulated in the presence of DNA. The ATPase activity in the presence of DNA was further stimulated by addition of the SopB protein. However, the SopB protein alone failed to stimulate the ATPase activity. 相似文献
105.
David J. Millar Antoni R. Slabas Chris Sidebottom Colin G. Smith Anthony K. Allen G. Paul Bolwell 《Planta》1992,187(2):176-184
A major wall protein of suspension-cultured cells of French bean has been isolated and characterised. It can be prepared from walls or the culture filtrate and in composition it is particularly rich in proline, valine and glutamic acid/glutamine and contains appreciable amounts of hydroxyproline. The N-terminus shows some glycosylation, while following chemical deglycosylation the first 38 residues were found to be identical to those of proline-rich proteins from soybean. However, the composition of the highly purified Mr-42000 bean protein differs considerably from the soybean proteins and must contain its own specific domains. An antibody was raised and used to demonstrate the inducibility of the Mr-42000 bean protein in response to elicitor action. The protein was found to be mainly localised in the intercellular spaces of the cortical cells of bean hypocotyls and at the wall-plasmalemma interface of xylem vessels, another potentially accessible compartment for pathogens. Following wounding, the protein was found to be generally distributed in the wall of epidermal and cortical cells of the hypocotyls. The Mr-42000 protein is cross reactive with antibodies raised to glycoproteins of the Rhizobium infection thread and the chitin-binding hydroxyproline-rich glycoprotein, potato lectin. These common epitopes together with the previously demonstrated chitin-binding properties of the bean protein indicate a role in host-microbial interactions. Furthermore, the Mr-42000 protein itself bound to the growing hyphal tips of the bean pathogen, Colletotrichum lindemuthianum.Abbreviations FITC
fluorescein isothiocyanate
- IgG
immunoglobulin G
- PAL
phenylalanine ammonia-lyase
We thank Dr Nick Brewin for advice on interpretation of immunolocalisations and for the gift of MCA 265. We thank Dudley Fernandino for carrying out the confocal microscopy. GPB thanks the Science and Engineering Research Council for funding. 相似文献
106.
cDNA cloning of an extracellular dermal glycoprotein of carrot and its expression in response to wounding 总被引:2,自引:0,他引:2
Suspension-cultured cells of carrot (Daucus carota L.) synthesize and secrete a glycoprotein that is normally found only in dermal tissues (epidermis, endodermis and periderm). This protein, previously called GP57, is now referred to as EDGP (E xtracellular D ermal G lyco P rotein). We purified sufficient quantities of EDGP to obtain amino-acid sequences on two internal tryptic peptides and screened a cDNA library of young carrot roots with antiserum to EDGP and with oligonucleotides corresponding to the peptides. Here we report the derived amino-acid sequence of EDGP. Sequence comparisons show that it has 40% amino-acid sequence identity with 7S basic globulin, a protein that is released when soybean seeds are soaked in hot water for a few hours. We suggest that these two proteins belong to a new family of dermal proteins. As far as we know, this is the first reported derived amino-acid sequence for protein that is specific to the epidermis and other dermal tissues. The level of EDGP mRNA is low in dry seeds, but increases rapidly in growing seedlings as they develop dermal tissues. The level of mRNA is low in storage roots, but increases rapidly in response to wounding. The presence of EDGP in dermal tissues and its up-regulation in response to wounding indicate a role in the response of plants to biotic and-or abiotic stresses. An unusual feature of the amino-acid sequence of EDGP is that it contains a short motif, which is present at the active site of aspartyl proteases such as pepsin and chymosin.Abbreviations cDNA
copy DNA
- 2,4-D
2,4-dichlorophen-oxyacetic acid
- EDGP
extracellular dermal glycoprotein
- 7SBG
7S basic globulin
Supported by a contract from the United States Department of Energy (Energy Biosciences) (to M.J.C.) and a Grant-in-Aid for Special Research on Priority Areas (01660002, Cellular and Molecular Basis for Reproductive Processes in Plants) from the Ministry of Education, Science and Culture, and by the Fund from Basic Research Core System of Science and Technology Agency, Japan (to S.S.). 相似文献
107.
We previously demonstrated that the oxysterol potentiation of arachidonic acid release and prostaglandin biosynthesis induced by foetal calf serum activation of normal rat kidney (NRK) cells (fibroblastic clone 49F) was not related to a direct effect of oxysterols on cell free Ca2+ level. Since both Ca2+ variations and protein C are involved in arachidonic acid release in some models, we looked for a possible modulation by protein C in the oxysterol effect on arachidonic acid release. We show that when the phorbol ester 12-O-tetradecanoyl-phorbol-13acetate (TPA), a protein kinase C activator, was added to the culture medium, the oxyterol effect on arachidonic acid release and prostaglandin synthesis clearly increased. Moreover, the effect of TPA was dose-dependent and TPA EC50 (4 × 10−9 M) was unchanged in the presence of the oxysterol. Preincubation of cells with TPA for 24 h prevented the arachidonic acid release induced by TPA alone, whereas the oxysterol effect was decreased but not abolished. In the absence of serum, TPA and ionomycin added together induced the same noticeable (arachidonic acid) release and PGE2 synthesis as serum alone. Nevertheless, the potentiating effect of cholest-5-ene-3β,25-diol was much higher when serum itself was used to activate NRK cells than it was in the present serum-mimicking experimental conditions. Thus, the presence of growth factors is probably required to obtain a full oxysterol effect. We conclude that the oxysterol effect was synergistic with, but not fully dependent on, protein kinase C and Ca2+ ion fluxes, therefore oxysterols could affed earlier events triggered by serum growth factor binding to their cell membrane receptors. 相似文献
108.
Sachio Hayashi Masaharu Nonoguchi Yoshihiko Shimokawa Mikihiko Tubouchi Yoshiyuki Takasaki Kiyohisa Imada 《Journal of industrial microbiology & biotechnology》1992,9(2):145-147
Summary Two extracellular -fructofuranosidases (E-1 andE-2) fromAureobasidium sp. ATCC 20524, producing 1-kestose (1F--fructofuranosyl-sucrose) from sucrose, were purified to homogeneity. Molecular weights of the enzymes were estimated to be about 304000 (E-1) and 315000 (E-2) Da by gel filtration. The enzymes contained 33% (w/w) (E-1) and 27% (w/w) (E-2) carbohydrate. TheK
m values for sucrose ofE-1 andE-2 andE-2 were 0.34 and 0.28 M, respectively. were 0.34 and 0.28 M, respectively. The enzymatic profiles of these enzymes were almost identical to intracellular enzymesP-1 andP-2 except for the differences in carbohydrate content andK
m values ofE-2 andP-2. 相似文献
109.
Dr. Yoko Kameda 《Cell and tissue research》1982,225(3):693-697
Summary Small follicles composed solely of C cells were occasionally observed in large C cell groups of dog thyroid glands. The lumina of C-cell follicles were filled with, or contained peripheral depositions of PAS-positive amorphous material, which was similar in ultrastructural features to thyroglobulin-containing colloid in typical thyroid follicles. This indicates that C cells, in addition to secreting calcitonin, produce a glycoprotein that can be stored in the lumina of the follicles. 相似文献
110.
James C. Fuscoe J.Patrick ONeill Richard Machanoff Abraham W. Hsie 《Mutation research》1982,96(1):15-30
We describe an assay for the quantification of reverse mutations at the hypoxanthine-guanine phosphoribosyltransferase (hgprt) locus in Chinese hamster ovary cells utilizing the selective agent L-azaserine (AS). Conditions are defined in terms of optimal AS concentration, cell density, and phenotypic expression time. After treatment, replicate cultures of 106 cells are allowed a 48-h phenotypic expression time in 100-mm plates. AS (10 μM) is then added directly to the growing culture and AS-resistant (ASr) cells form visible colonies. This assay is used to quantify ICR-191-, ICR-170-, and N-ethyl-N-nitrosourea-induced reversion of independently isolated HGPRT? clones. The ASr phenotype is characterized both physiologically and biochemically. All ASr clones isolated are stably resistant to AS and aminopterin but sensitive to 6-thioguanine. They also have re-expressed HGPRT enzyme. In addition, several revertants are shown to contain altered HGPRT. The data provide further evidence that ICR-191 and ICR-170 cause structural gene mutations in mammalian cells and also suggest that ICR-191, ICR-170, and N-ethyl-N-nitrosourea induce similar types of mutations in Chinese hamster ovary cells. 相似文献