全文获取类型
收费全文 | 5538篇 |
免费 | 316篇 |
国内免费 | 1篇 |
专业分类
5855篇 |
出版年
2023年 | 15篇 |
2022年 | 21篇 |
2021年 | 67篇 |
2020年 | 46篇 |
2019年 | 62篇 |
2018年 | 73篇 |
2017年 | 78篇 |
2016年 | 119篇 |
2015年 | 166篇 |
2014年 | 196篇 |
2013年 | 297篇 |
2012年 | 546篇 |
2011年 | 1084篇 |
2010年 | 540篇 |
2009年 | 578篇 |
2008年 | 243篇 |
2007年 | 245篇 |
2006年 | 257篇 |
2005年 | 203篇 |
2004年 | 232篇 |
2003年 | 194篇 |
2002年 | 182篇 |
2001年 | 40篇 |
2000年 | 28篇 |
1999年 | 45篇 |
1998年 | 44篇 |
1997年 | 24篇 |
1996年 | 27篇 |
1995年 | 24篇 |
1994年 | 28篇 |
1993年 | 18篇 |
1992年 | 19篇 |
1991年 | 19篇 |
1990年 | 8篇 |
1989年 | 11篇 |
1988年 | 10篇 |
1986年 | 2篇 |
1985年 | 4篇 |
1984年 | 3篇 |
1983年 | 6篇 |
1968年 | 3篇 |
1967年 | 2篇 |
1962年 | 2篇 |
1874年 | 3篇 |
1873年 | 4篇 |
1872年 | 4篇 |
1871年 | 2篇 |
1858年 | 2篇 |
1856年 | 5篇 |
1855年 | 4篇 |
排序方式: 共有5855条查询结果,搜索用时 15 毫秒
21.
Martine Crasnier Valérie Dumay Antoine Danchin 《Molecular genetics and genomics : MGG》1994,243(4):409-416
In Escherichia coli, adenylate cyclase activity is regulated by phosphorylated EnzymeIIAGlc, a component of the phosphotransferase system for glucose transport. In strains deficient in EnzymeIIAGlc, CAMP levels are very low. Adenylate cyclase containing the D414N substitution produces a low level of cAMP and it has been proposed that D414 may be involved in the process leading to activation by EnzymeIIAGlc. In this work, spontaneous secondary mutants producing large amounts of cAMP in strains deficient in EnzymeIIAGlc were obtained. The secondary mutations were all deletions located in the cya gene around the D414N mutation, generating adenylate cyclases truncated at the carboxyl end. Among them, a 48 kDa protein (half the size of wild-type adenylate cyclase) was shown to produce ten times more cAMP than wild-type adenylate cyclase in strains deficient in EnzymeIIAGlc. In addition, this protein was not regulated in strains grown on glucose and diauxic growth was abolished. This allowed the definition of a catalytic domain that is not regulated by the phosphotransferase system and produces levels of cAMP similar to that of regulated wild-type adenylate cyclase in wild-type strains grown in the absence of glucose. Further analysis allowed the characterization of the COOH-terminal regulatory domain, which is proposed to be inhibitory to the activity of the catalytic domain. 相似文献
22.
John S. Beck Anne E. Kwitek Phillip H. Cogen Andrew K. Metzger Geoffrey M. Duyk Val C. Sheffield 《Human genetics》1993,91(1):25-30
p53 is a tumor suppressor gene located on 17p, a region of the human genome frequently deleted in tumors. Mutation of the p53 gene is an important step leading to development of many forms of human cancer. To simplify the analysis of tumors for p53 point mutations, we describe a GC-clamped denaturing gradient gel assay for detecting single-base substitutions within highly conserved regions of the p53 gene. This assay alows for efficient screening of tumors for single-base substitutions within the p53 gene and can be used to facilitate sequence analysis of p53 point mutations. 相似文献
23.
Valérie Michel Gérard Fonty Liliane Millet Frédérique Bonnemoy Philippe Gouet 《FEMS microbiology letters》1993,110(1):5-9
Abstract To better define the antigenic structure of the outer cell membranes of Legionellae, a panel of 6 monoclonal antibodies was raised against partially purified outer membranes of Legionella pneumophila serogroup 1, Corby strain. This study describes the purification and characterization of one of these monoclonal antibodies reacting with a 135-kDa protein, which was shown to be common to all 14 serogroups of Legionella pneumophila . It shows no cross-reactivity with 20 other Legionella species, or 9 other Gram-negative species tested by SDS-PAGE and Western blotting procedures. The epitope would appear to be predominantly surface exposed and, from preliminary detergent extraction studies, not peptidoglycan-associated. 相似文献
24.
Valéria Marques Gabriel Riaño Miguel A. Carretero Iolanda Silva-Rocha Catarina Rato 《Acta zoologica》2023,104(3):419-433
Under temperature sex determination (TSD), sex is determined by temperature during embryonic development. Depending on ecological and physiological traits and plasticity, TSD species may face demographic collapse due to climate change. In this context, asymmetry in bilateral organisms can be used as a proxy for developmental instability and, therefore, deviations from optimal incubation conditions. Using Tarentola mauritanica gecko as a model, this study aimed first to confirm TSD, its pattern and pivotal temperature, and second to assess the local adaptation of TSD and variation of asymmetry patterns across four populations under different thermal regimes. Eggs were incubated at different temperatures, and hatchlings were sexed and measured. The number of lamellae was counted in adults and hatchlings. Results were compatible with a TSD pattern with males generated at low and females at high incubation temperatures. Estimated pivotal temperature coincided with the temperature producing lower embryonic mortality, evidencing selection towards balanced sex ratios. The temperature of oviposition was conservatively selected by gravid females. Asymmetry patterns found were likely related to nest temperature fluctuations. Overall, the rigidity of TSD may compromise reproductive success, and demographic stability in this species in case thermal nest choice becomes constrained by climate change. 相似文献
25.
Valérie Copié John A. Battles John M. Schwab Dennis A. Torchia 《Journal of biomolecular NMR》1996,7(4):335-340
Summary Nearly complete backbone 1H, 15N and 13C signal assignments are reported for -hydroxydecanoyl thiol ester dehydrase, a 39-kDa homodimer containing 342 amino acids. Although 15N relaxation data show that the protein has a rotational correlation time of 18 ns, assignments were derived from triple-resonance experiments recorded at 500 MHz and pH 6.8, without deuteration. The Chemical Shift Index, CSI, identified two long helices and numerous -strands in dehydrase. The CSI predictions are in close agreement with the secondary structure identified in the recently derived crystal structure, particularly when one takes account of the numerous bulges in the -strands. The assignment of dehydrase and a large deuterated protein [Yamazaki et al. (1994) J. Am. Chem. Soc., 116, 11655–11666] suggest that assignment of 40–60 kDa proteins is feasible. Hence, further progress in understanding the chemical shift/structure relationship could open the way to determine the structures of such large proteins.
Supplementary Material is available on request, comprising Table S1 listing the spectral parameters; Table S2 listing the assignments; Fig. S1 showing the 2D 1H–15N HSQC spectrum; Fig. S2 showing sequential NOEs, secondary shifts, H-exchange and 3JHN
data; and Fig. S3 showing plots of the H, C, CO and C Chemical Shift Indexes.To whom correspondence should be addressed. 相似文献
26.
Rachel Baltz Jean-Luc Evrard Val/'erie Bourdon Andr/'e Steinmetz 《Sexual plant reproduction》1996,9(5):264-268
The protein PLIM-1 (formerly SF3) from sunflower is expressed exclusively in mature, free pollen. It contains two LIM domains associated with an acidic C-terminus comprising six copies of the pentapeptide motif (A,T,S) (E,D) TQN. We have expressed the pollen protein as well as some of its mutant forms inEscherichia coli and have used the bacterially produced proteins to study interactions with nucleic acids. Our studies show that the protein binds DNA and RNA in vitro to form large complexes, while mutant polypeptides containing either a single LIM domain or a destabilized first or second LIM domain do not. Although these data suggest that the biological function of PLIM-1 involves interactions with nucleic acids, its role in pollen development remains unclear. 相似文献
27.
The hydrological structure of the French coastal part of the eastern English Channel is strongly linked with tidal regimes and riverine input. Two distinct water masses are separated by a frontal area and drift along the coast in SW–NE direction. These two water masses are well-mixed during the entire year. We studied the seasonal dynamic of nitrogenous nutrients, chlorophyll a and organic particulate carbon and nitrogen at two stations, characteristic of these water masses, during the year 1994. Results show (i) a winter stock of nitrate and ammonium, (ii) a pre-bloom period corresponding to the use of ammonium, (iii) a high bloom period of short duration using nitrate, (iv) a post-bloom period with little phytoplanktonic activity probably limited by nutrients other than nitrogen and (v) an autumnal period of reconstitution of stock. The essential difference between the two stations is the importance of winter stock of nutrients and of bloom chlorophyll a concentration, with the coastal station richer than the offshore one. An assumption about the nitrogen available for new production in this area gives a value of 57% of the winter stock of inorganic nitrogen. 相似文献
28.
Emmanuel Martin Valère Cacheux Hélène Cavé Jean Michel Lapierre Denis Le Paslier Bernard Grandchamp 《Human genetics》1995,96(6):668-670
CDKN4/p27Kip1 is a cyclin-dependent kinase (Cdk) inhibitor implicated in G1 phase arrest, which negatively regulates G1 phase progression in response to TGF, and might represent a tumor suppressor gene. We report here the chromosomal assignment of the human CDKN4 gene to chromosome 12p12.3 in close proximity to highly polymorphic microsatellite markers. 相似文献
29.
Effects of Physicochemical Factors on the Adhesion to Cellulose Avicel of the Ruminal Bacteria Ruminococcus flavefaciens and Fibrobacter succinogenes subsp. succinogenes
下载免费PDF全文
![点击此处可从《Applied microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Valrie Roger Grard Fonty Sylvie Komisarczuk-Bony Philippe Gouet 《Applied microbiology》1990,56(10):3081-3087
Ruminococcus flavefaciens adhered instantly to cellulose, while Fibrobacter succinogenes had the highest percentage of adherent cells after about 25 min of contact between bacteria and cellulose. Adhesion of R. flavefaciens was unaffected by high concentrations of sugars (5%), temperature, pH, oxygen, metabolic inhibitors, and lack of Na+. In contrast, the attachment was affected by the removal of divalent cations (Mg2+ and Ca2+), the presence of cellulose derivatives (methylcellulose and hydroxyethylcellulose), and cystine. Adhesion of F. succinogenes was sensitive to low and high temperatures, high concentrations of glucose and cellobiose (5%), hydroxyethylcellulose (0.1%), redox potential, pH, lack of monovalent cations, and the presence of an inhibitor of membrane ATPases or lasalocid and monensin. Cells of F. succinogenes heated at 100°C no longer were adherent. On the other hand, adhesion was insensitive to the lack of divalent cations (Mg2+ and Ca2+), the presence of 2,4-dinitrophenol, tetrachlorosalicylanilide, or inhibitors of the electron transfer chains. Adhesion of F. succinogenes seems to be related to the metabolic functions of the cell. External proteins and/or cellulases themselves might play a part in the attachment process. Several mechanisms are probably involved in the adhesion of R. flavefaciens, the main one being the interaction between the large glycocalyx and the divalent cations Ca2+ and Mg2+. Hydrophobic bonds and enzymes may also be involved. 相似文献
30.
Pyruvate decarboxylase (PDC) was prepared from four-day-old pea seedlings by a procedure which involves extraction of plant
material with a phosphate buffer, fractionation of the extract with ammonium sulphate, desalting by dialysis or gel filtration
on Sephadex G-25 column, chromatography on DEAE cellulose and filtration on Sepharose 4B. The PDC preparation activity 10
000 U g-1 protein was about 600 fold higher than that of the sodium phosphate buffer extract. According to the enzyme behaviour during
the gel filtration on different carriers the molecular mass of pea PDS was estimated at about 106. Magnesium ions and thiamine pyrophosphate were found to be coenzymes of PDC. Cofactors can be removed by 48 h dialysis followed
by chromatography on DEAE cellulose. Apoenzym is activated optimally with the concentration of cofactors of 0.002 M. Magnesium
ions can be replaced in their activation function by ions of Fe2+, Ni2+, Co2+, Zn2+ and Mn2+. Another ions,i.e. Ba2+, Ca2+, Cd2+, Hg2+ and Cu2+ are inactive. Assumption about the relation between the ion diameter and degree of activation is formulated. 相似文献